National Algal Biofuels
Technology Roadmap

MAY 2010

National Algal Biofuels
Technology Roadmap
A technology roadmap resulting from the National Algal Biofuels Workshop
December 9-10, 2008
College Park, Maryland

Workshop and Roadmap sponsored by the U.S. Department of Energy
Office of Energy Efficiency and Renewable Energy
Office of the Biomass Program
Publication Date: May 2010

John Ferrell Valerie Sarisky-Reed
Office of Energy Efficiency Office of Energy Efficiency
and Renewable Energy and Renewable Energy
Office of the Biomass Program Office of the Biomass Program
(202)586-5340 (202)586-5340

Roadmap Editors:
Daniel Fishman, Rajita Majumdar,1 Joanne Morello,2 Ron Pate,3 and Joyce Yang2

Workshop Organizers:
Al Darzins, Grant Heffelfinger, Ron Pate, Leslie Pezzullo,2 Phil Pienkos,4 Kathy Roach,5 Valerie Sarisky-Reed,2 and the
4 3 3

Oak Ridge Institute for Science and Education (ORISE)

A complete list of workshop participants and roadmap contributors is available in the appendix.

Suggested Citation for This Roadmap:
U.S. DOE 2010. National Algal Biofuels Technology Roadmap. U.S. Department of Energy, Office of Energy Efficiency and
Renewable Energy, Biomass Program.
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BCS, Incorporated 2Office of the Biomass Program 3Sandia National Laboratories

National Renewable Energy Laboratory 5MurphyTate LLC

This report is being disseminated by the Department of Energy.2 of the Bulletin. . As such. This report has been peer reviewed pursuant to section II. this report could be “influential scientific information” as that term is defined in the Office of Management and Budget’s Information Quality Bulletin for Peer Review (Bulletin). 106-554) and information quality guidelines issued by the Department of energy. Further. the document was prepared in compliance with Section 515 of the Treasury and General Government Appropriations Act for Fiscal Year 2001 (Public Law No.

Renewable energy lies at the intersection of some of our nation’s most pressing issues: our energy security. Meanwhile. comments gathered during a public comment period. It is intended to serve as a resource for researchers. and decision-makers by providing a summary of progress to date and a direction for future algae RD&D activities. research managers. and the stability of our global climate. Through appropriated dollars and the American Recovery and Reinvestment Act of 2009.S. Valerie Sarisky-Reed Acting Manager. engineers. and other stakeholders. Achieving cost-competitive. it remains committed to fostering technologies that could yield substantial benefits over time. and supporting scientific literature. and the academic community are also increasing their efforts to optimize and commercialize this renewable energy source. our economic wellbeing. Prepared with the input of more than 200 scientists. Department of Energy Preface  i . This Roadmap lays the groundwork for identifying challenges that will likely need to be surmounted for algae and cyanobacteria to be used in the production of economically viable. engineers. this document represents the synthesis of the Biomass Program’s National Algal Biofuels Technology Roadmap Workshop. DOE looks forward to continuing its work with diverse partners in evaluating renewable energy options and facilitating development of those that carry the greatest benefits today and in the years to come.S. while DOE works to deploy renewable energy and energy-efficient projects across the country today. but their significant potential to serve as renewable transportation fuels warrants our thorough consideration of what efforts are necessary to make them a commercial-scale reality. Biomass Program U. sustainable algal biofuels will entail years of research and careful planning. reliable energy. and we hope it fosters and informs participation from existing and new stakeholders as the next steps are taken to advance algal biofuels. as well as the recognition that we will not likely achieve these goals via one technology pathway. Department of Energy (DOE) Biomass Program’s National Algal Biofuels Technology Roadmap. industry representatives. and demonstration (RD&D) projects that seek to tackle key technical hurdles associated with commercializing algal biofuels. DOE is investing in a variety of research. DOE has recently revived its investment in algal biofuels in response to the increased urgency of lowering greenhouse gas emissions and producing affordable. environmentally sound biofuels.Preface Welcome to the U. other federal agencies. development. This Roadmap is intended to summarize the state of technology for algae-based fuels and document the research and development challenges associated with producing them at a commercial scale. These national challenges require near-term solutions as well as investments in nascent technologies that show promise for the future. Therefore. private companies.

brackish. creates economic and social uncertainties for businesses and individuals. ii Executive Summary  . meet the new RFS feedstock resources. Much has changed since deployment of commercial algae-to-biofuel processes. weight). which exposes the country to the risk of critical disruptions in fuel supply. fungible Workshop. a number of next generation have combined to capture the interest of researchers and biofuels show significant promise in helping to achieve entrepreneurs around the world. illustrated the potential Program to invest in the research.S. cellulosic biofuels and biomass-based diesel. Rising petroleum prices and a Additional funding is being directed to algae-to-biofuel national mandate to reduce U. and macroalgae (or While the basic concept of using algae as an alternative seaweed).Executive Summary “ ” Developing the next generation of biofuels is key to our effort to end our dependence on foreign oil and address the climate crisis – while creating millions of new jobs that can’t be outsourced — Secretary of Energy Steven Chu at the White House ceremony on May 5. Of these candidates. including starches and brought together more than 200 scientists. This announcement included funds for the Department of Energy (DOE) Office of Energy The DOE-supported Aquatic Species Program. These include: the goal. produced. marine. some microalgae and renewable source of biomass feedstock for biofuels have the potential to accumulate significant amounts has been explored previously. Renewable Fuel Standard (RFS) requiring transportation sugars from cyanobacteria and macroalgae. a scalable.S. the end of the program. 5) production of both biofuels and valuable Energy Steven Chu announced the investment of $800M co-products. and Renewal Act. including advanced and products can be generated using algae precursors. and create economic and programs. These fuels can also be produced using other algae by DOE-EERE’s Biomass Program. 2009. and impacts A culture of the microalgae Botryococcus. sustainable of lipids (more than 50% of their ash-free cell dry and commercially viable system has yet to emerge. While cellulosic ethanol is expected to play a large role There are several aspects of algal biofuel production that in meeting the EISA goals. President Obama and Secretary of wastewater). opportunities across the nation have renewed interest in developing algal feedstocks for biofuels production. 2008. and 6) potential recycling of CO2 and other in new research on biofuels in the American Recovery nutrient waste streams. In addition fuel sold in the U. dependence on foreign research both in EERE and other government agencies oil. The term algae can refer to microalgae. to contain a minimum of 36 billion to fungible biofuels. cyanobacteria (the so called “blue-green algae”). 4) utilization of a wide variety of water energy independence. biomass-derived fuels have received increasing attention as one solution to our nation’s continued and growing dependence on imported oil. saline. was convened fuels. Photo courtesy of the our national security. engineers. and advanced biofuels. The two-day event feedstocks and intermediates. 3) use of otherwise non-productive. while at the same time moving the nation ever closer to non-arable land. biofuels derived from algae 1) high per-acre productivity. To accelerate the deployment of sources (fresh. announcing $800 million in new biofuel research activities I n recent years. by 2022. These characteristics give great potential for The National Algal Biofuels Technology Roadmap an immediate pathway to high energy density. The Energy Independence and Provasoli-Guillard National Center for Culture of Marine Phytoplankton Security Act of 2007 (EISA) established a mandatory (CCMP). 2) non-food based have the potential to help the U. and of algae as a biofuel feedstock.S. provide environmental benefits. development. Under certain conditions. held December 9-10. a variety of different biofuels and gallons of renewable fuels. an effort Efficiency and Renewable Energy’s (EERE) Biomass undertaken from 1978 to 1996.

the Roadmap Workshop effort suggests document is intended to provide a comprehensive state that many years of both basic and applied science and of technology summary for fuels and co-products from engineering will likely be needed to achieve affordable. guide researchers and engineers. industry representatives. Executive Summary  iii . development. By documenting the challenges across the algal discuss and identify the critical challenges currently biomass supply chain and highlighting research and hindering the economical production of algal biofuels at coordination needs and gaps. policymakers. this document will serve to commercial scale.research managers. research. and regulators from across the country to scale. This document also seeks to explore technologies in an integrated process will be a key the economic and environmental impacts of deploying component to accelerating progress. lawyers. and sustainable algal-based fuels. and the private sector in implementing national This document represents the output from the Workshop. algal biomass production systems at commercial financiers. The Roadmap In summary. and comments received during a public comment period. federal agencies. and deployment efforts. The ability techno-economic challenges associated with scaling to quickly test and implement new and innovative up of processes. algal feedstocks and to document the feasibility and scalable. supporting scientific literature.

FROM ALGAE FROM TO TO ALGAE BIOFUELS BIOFUELS An Integrated Systems An Integrated Approach Systems to Renewable Approach Energy to Renewable that is E nergy ALGAE FEEDSTOCKS CULTIVATION Microalgae and cyanobacteria can be cultivated via photoautotrophic methods (where algae require light to grow and create new biomass) in open or closed ponds or via heterotrophic methods (where algae are grown without light and are fed a carbon source. Macroalgae (or seaweed) has different cultivation needs that typically require open off-shore or coastal facilities. Understanding. and taking advantage of the biology of algal strains selected for use in production systems is the foundation for processing feedstocks into fuels and products. macroalgae (seaweed). to generate new biomass). and cyanobacteria (formerly called “blue-green algae”). Isolating new strains directly from unique environments will ensure versatile and robust strains for mass culture needed in biofuels applications. Algae occur in a variety of natural aqueous and terrestial habitats ranging from freshwater. such as sugars. and hyper-saline environments to soil and in symbiotic associations with other organisms. Open Ponds Example Cultivation Systems POLICY SITING AND RESOURCES iv Executive Summary  Development Systems and Techno-Economic Analysis: Guiding Path Toward the Research and a . brackish waters. managing. Designing an optimum cultivation system involves leveraging the biology of the algal strain used and inegrating it with the best suited downstream processing options. marine. scalability. and sustainability of algae to biofuel systems. Fermentation Tanks MICROALGAE CYANOBACTERIA MACROALGAE Algae as feedstocks for bioenergy refers to a diverse group of Closed Photobioreactors organisms that include microalgae. Choices made for the cultivation system are key to the affordability.

g. and proteins. issues. Algal cultures are mainly grown 1) Conversion of whole algal biomass. industrial enzymes. Focusing on biofuels as the end-product O poses challenges due to the high volumes and relative low CH2-O-C R1 values associated with bulk commodities like gasoline and O diesel fuels. gasoline. optimizing extraction systems that consume less energy than contained in the algal products is a challenge due to the high energy needs associated with both handling and drying algal biomass as well as separating out desirable products.g. or a 2-O-C EXTRACTION combination of these approaches. These steps can be energy-intensive and can entail siting Conversion technology options include chemical. carbohydrates. The end products vary depending on the conversion technology utilized. CH-O-C R2 O CH2-O-C R3 Bio-Crude Algal Lipid: Precursor to Biofuels Three major components can be extracted from algal biomass: lipids (including triglycerides and fatty acids). End Uses: • Biodiesel • Biogas • Renewable Hydrocarbons • Co-products • Alcohols (e.. and thermochemical processes.. and Sustainable HARVESTING / DEWATERING CONVERSION Some processes for the conversion of algae to liquid Conversion to fuels and products is predicated on a basic transportation fuels require pre-processing steps such as process decision point: harvesting and dewatering.g. animal feed. and surfactants) REGULATIONS AND STANDARDS Executive Summary  v Commercially Viable Algal Biofuel Industry . biochemical. biodiesel and jet fuel).. Abundant. though these are also subject to a number of unique scale-up challenges. animal/fish feeds). Affordable. Some algal biomass production processes are investigating options to bypass extraction. bioplastics. While many analytical techniques exist. fertilizers. Most challenges in extraction are associated with the industrial scale up of integrated extraction systems. While lipids and carbohydrates are fuel precursors (e. in water and can require process steps to concentrate 2) Extraction of algal metabolites. proteins can be used for co-products (e. or harvested algal biomass prior to extraction and conversion. 3) Processing of direct algal secretions.

vi Executive Summary  .

........................................................................20 Considerations of Genetic Modifications........................ 31 Contents  vii ................................................................................................ 16 The Genetic Toolbox.......9 Selecting Algal Model Systems for Study.................................................................................................................................. Algal Cultivation...........................3 1........................... 23 3........................................................................................................ Contents 1.........................2 Scale-Up Challenges.................................................................. Overview............................................................................................................ 29 Microalgae and Cyanobacteria............................................................................. 11 Photosynthesis and Light Utilization.............................................................................................................................................................................................................. 12 Lipid Synthesis and Regulation......................................................................................................................................5 1...........................................3 Algal Biotechnology...............................................................................................................................................................3 Early Work to 1996..................................................... 15 2........................................................................................................................................................................................................................................................................................................................................................................................................................................................................2 Algal Physiology and Biochemistry.............................................................................. ............................................................................................8 2.............................................................................................................................................................................................1 About the Roadmap......30 3......................................... 31 Stability of Large-Scale Cultures .................................10 2.........8 Role of Culture Collections as National Algae Data Resource Centers.................7 References...... 12 Biohydrogen .......................... Screening and Selection .........................................................................3 Algae-to-Biofuels: Opportunity and Challenges Today......5 Regulations and Standards.........................................................................................................29 3...........................................................................7 2................................................................................................................................................................................................................................... 22 References............................................... 19 Applications of Biotechnology to Algal Bioenergy......................................................................................................................................................................................................8 Isolation and Characterization of Naturally Occurring Algae ..................1 Advantages of Algal Feedstocks..............................................................................................................................................................................................................................................................................5 Technology and Analysis Challenges.......1 Strain Isolation..................................1 America’s Energy Challenges.............................................................7 Public-Private Partnerships......................................11 Carbon Partitioning and Metabolism.................. Algal Biology..............................................................................................................................................................................................................................2 A History of Domestic Algal Biofuels Development..........3 Research from 1996 to Present.............................11 Algal Carbohydrates..........................1 1..........................................................................................................................1 Cultivation Pathways........... 16 Enabling Technologies: “Omics” Approaches and Bioinformatics........................................................................................................................ 29 Macroalgae......................................................................................................

...................................................... .......................................... 38 Harvesting...................................................................................................................37 Harvesting..................................................................................................................................................................................2 Processing of Whole Algae...................................40 Preliminary Look at Energy Balance ...44 Selective Extraction...............1 Direct Production of Biofuels from Algae..............................................................48 Alcohols................................... System Productivity....................................................................................................................................................................................................................................................................................................................................44 Subcritical Water Extraction................................................50 6............................................................................................................................................................2 Challenges...........................................46 6.............................40 5..................................................... 51 Liquefaction....................................................................42 5............................................................................................48 6................................................................................................................................................................................................................................................................................... 49 Alkanes.......................................................................................................................................................................................................................................................................................................... 52 Supercritical Processing.......................................... 37 4.............................................................................. and Management................................................................................................................................................................................................................................................................................................................. 39 References..................... Cell Rupture)......................................................................................................................................................................................................................................................................................................................... Sustainability........................ 53 Anaerobic Digestion of Whole Algae....................................................................................................................................................................................................................... 35 4.................................................................... 45 Supercritical Fluid Extraction..................................................... 33 References....................................................46 References................................................ 32 Nutrient Sources..................e............................ and Recycling..................................................3 Systems Engineering ...............................................50 Gasification.......................................................................................................... 45 Heterotrophic Production.......................................................................................................................................................................... 32 Water Management.............................................. ................. Downstream Processing: Harvesting and Dewatering............. 49 Hydrogen..................................3 Conversion of Algal Extracts..............50 Pyrolysis............................................................ Conservation..... 42 Mechanical Disruption (i.............................................................................................................................. Algal Biofuel Conversion Technologies .................................................. 38 4...........................................................................................................2 Approaches for Macroalgae ...............................................................1 Approaches for Microalgae.......................... 53 6.......................1 Current Practices for Lipid Extraction...........................37 Drying..................... 42 Organic Co-solvent Mixtures................................................................................................................. 43 Accelerated Solvent Extraction.. 54 viii Contents  ................................................................................................................ 39 4................................................................................................................................... 45 5........................ 38 Preprocessing.................................................................................................... 53 Chemical Transesterification.......................................................................................................................................................................................................................................... Extraction of Products from Algae .......................................

....................................................................................................................................................................2 Commercial Products from Macroalgae................................74 9.............................................. Resources and Siting .......................................................................................................4 Processing of Algal Remnants after Extraction.......... 84 Main Research Needs for Algae Production with Wastewater .................................................64 7......................................................................................................................................................................................................................................................................................................73 Heterotrophic Microalgae Approach................................................................................................. 87 Barriers to Co-Location of Algae Production with Stationary Industrial CO2 Sources ............................................ 58 7..............................................................................................................................................................................................................................................................................4 Integration with Water Treatment Facilities .........................5 Co-location of Algal Cultivation Facilities with CO2-Emitting Industries................................................................................. 73 Photoautotrophic Microalgae Approach................................................................57 References............................................................................................................................................................................................................................................................................ 55 Conversion to Renewable Diesel..........................71 References........80 Land......64 References.................................. ........................................................................................75 Water......................................................................................... Biochemical (Enzymatic) Conversion.............................. Co-products ......................................................................................................... Distribution and Utilization....... 61 7........ 88 Directions for Research and Development............................................................................... 86 The Opportunity in Co-Locating with CO2-Emitting Sources...... 70 Algal Blendstocks for Alcohol and Gasoline-Range Petroleum Products.................. and Jet Fuel ....................................... ........................... 83 Algae Production Techniques for Wastewater Treatment Plants...........2 Resources Overview......................... 55 Catalytic Cracking.................................................77 Carbon Dioxide...........................................................................................3 Potential Options for the Recovery of Co-products.................................... 81 9...................................................................................69 8......................................1 Commercial Products from Microalgae and Cyanobacteria.......................................................................................1 Resource Requirements for Different Cultivation Approaches................................. 73 9......... Gasoline........................................................................................................................................................................................................................................................................... 69 8..........................................................2 Utilization.................. 61 7.... 88 References.......................................................... 70 Algal Blendstocks to Replace Middle-Distillate Petroleum Products...............................................72 9.....................................................................................................................................1 Distribution...................................................75 Climate .......................................................................... 56 6........................74 Photoautotrophic Macroalgae Approach.................. 86 9.............................................................................. 67 8..........................................................................................................................................................................................................................................................90 Contents  ix ... 83 Wastewater Treatment and Recycling Applications...................

..................................................... 93 10............................. Systems and Techno-Economic Analysis............................................................ 105 11..........................................................................................................................................................................................................................................3 Suggestions for TEA and LCA Approach (Workshop Results and Discussion).......................................................... 111 References...........................................................2 Components of Successful Public-Private Partnerships.......99 Overview................................................... 97 10................................................................................................................114 Appendices ........ 104 10.........122 Appendix 3: Respondents to Request for Information on the Draft Roadmap.......................................... 109 11.......................................................................6 Preliminary System Dynamics Modeling .............................................................. 109 11....................................................10........................ 100 Impact of Geographic Variability of Inputs on Algal Biofuel Production Costs......................................................4 Systems Analysis.........115 Appendix 2: Contributors...............................................................................................................1 The Benefits of Algal Biofuels Public-Private Partnerships.93 10.............5 Modeling Partnerships for Technology Development..................................................................110 11.....................................................................................................................................................1 Objectives of Techno-Economic Analysis ........124 x Contents  ................................ Public-Private Partnerships ............................ 111 11.......99 Engineering Analyses and GIS Assessments....102 Life Cycle Analysis..............................................5 Algae Production Costs and Uncertainties.................2 TEA Scope and Methods.................................................................................................................110 11..................................................... 115 Appendix 1 : Workshop Participants..................................................................3 Forums for Public-Private Partnerships...........................................................................................................................................7 Potential Directions for R&D Effort ...............4 Partnership Challenges in the Algal Biofuels Industry.................................... 104 10............................................................................................................................ 104 10........................................................................................................................................................................................94 10............

60% of 2008. Department experience and expertise during a series of technical of Energy (DOE) Office of Energy Efficiency and break-out sessions that spanned critical aspects of the Renewable Energy is committed to advancing the vision algal biomass supply chain and cross-cutting issues. The The legislation establishes production requirements for Workshop provided a stimulating environment to explore domestic alternative fuels under the Renewable Fuels topics affecting the development of algal biofuels industry. 1. sustainable domestic biomass industry that The outcomes from the Workshop provided key inputs produces renewable biofuels. Workshop participants to evaluate the Roadmap for and deployment (RD&D) activities. and non-governmental organizations. academia. representing a diverse range of expertise from promote the use of renewable fuels. Overview  1 . Standard (RFS) that increase over time (Exhibit 1. in partnership fidelity and incorporate new information. the national laboratories. In doing so. which set new standards for More than 200 stakeholders were convened at the vehicle fuel economy. of which only 15 economic. and policy perspectives that can support billion gallons can be produced from corn-based ethanol. Furthermore. the Roadmap describes fuels. and creates be found at http://www. provides environmental benefits.1 America’s Energy Challenges About the Roadmap As global petroleum supplies diminish.orau. approximately two-thirds of its petroleum. development. and criticisms not captured during the Workshop. The United States currently imports Technology Roadmap Workshop.S. A major objective of the reserves. energy supply for transportation liquid transportation fuels. industry. the state of technology the current status of algae RD&D. All comments are noted in the Appendix of this document. held December 9-10. as well as made provisions that Workshop. Following the release of the initial draft of the Roadmap. energy efficiency. The Workshop was organized by the Biomass Program to rising energy demand in many rapidly developing discuss and identify the critical challenges currently countries around the world is beginning to create hindering the development of a domestic. Renewable Fuels Standard (RFS) mandate established within EISA. commercial. government and new energy technology research and development. gallons of renewable fuels by 2022. bioproducts and biopower. agencies. that both assesses the current state of technology and provides direction to the Program’s RD&D efforts. with national laboratories. it lays for producing algal biofuels is regarded by many in the groundwork for identifying challenges that likely the field to be in its infancy and there is a considerable need to be overcome for algal biomass to be used in amount of RD&D is needed to achieve affordable. the production of economically viable biofuels. and guide RD&D investment in algal biofuels. The which is used for producing transportation fuels. Overview The Workshop was able to capture the participants’ The Biomass Program (Program) of the U. the United The framework for National Algal Biofuels Technology States is becoming increasingly dependent upon foreign Roadmap was constructed at the Algal Biofuels sources of crude oil. academia. energy security. at the University of Maryland-College Park.S. Algal biofuels offer great promise in contributing to Every attempt was made to ensure that the Roadmap the Program’s vision. While Biofuels derived from algae can meet these l onger-term addressing the potential economic and environmental needs of the RFS and represent a significant opportunity benefits of using algal biomass for the production of to impact the U.S. reduces our dependence Roadmap. the Energy Independence and Security Act (EISA) was enacted. 1. economic opportunities across the the combustion of petroleum- Workshop was to gather the necessary information based fuels has created serious concerns about climate to produce an algal biofuels technology roadmap change from the greenhouse gas (GHG) emissions. including the Energy Independence and Security Act a 60-day public comment period was held to allow of 2007. the Program has undertaken a diverse portfolio of research. The RFS mandates blending of 36 billion This Roadmap presents information from a scientific. as well as helping to meet the development process was transparent and inclusive. intense competition for the world’s dwindling petroleum scale algal biofuels industry. to the development of this Algal Biofuels Technology enhances U.1). and sustainable algal-based biofuels. scalable. To accomplish its goals. In 2007. and industry.1. viewpoints. of a viable. The Program’s goals are driven by various federal policies and laws. Despite their potential. The full proceedings of the Workshop can on oil.

2 1.0 36.2). For example. oleaginous Exhibit 1. As required by EISA.0 30.5 1.0 a 5. advanced biofuels must demonstrate GHG emissions UNIQUE ADVANTAGES OF ALGAL FEEDSTOCK FOR across their life cycle that are at least 50% less than GHG ADVANCED BIOFUELS emissions produced by petroleum-based transportation • High area productivity fuels.5 a 18.15 2015 3.5 20.5 a 13.25 a 7.80 1. but no less than 1..0 2.0 a 11. However.35 13.5 a 9. 2009).0 26.95 2011 0.55 2014 1. Cellulosic feedstocks were • Recycles stationary emissions of carbon dioxide identified by the Billion Ton Study as a significant source • Compatible with integrated production of fuels and of biomass (Perlack et al. and approximately 5 times that of oil palm fuels for aviation and ground transport.75 16.. Potential oil yields from certain algae production of biofuels and bioproducts from components strains are projected to be at least 60 times higher than of biomass.0 24. 2005).6 11.0 a 21.5 0.95 12.0 2022 16.0 2019 8.25 2017 5.2 2013 1.0 15.1 Renewable Fuel Standard volume requirements (billion gallons) Cellulosic biofuels and biomass-based diesel are included in the advanced biofuel requirement.0 billion gallons.Advanced biofuels face significant challenges in meeting the ambitious targets set by EISA. Algal biomass may per acre of land on an annual basis (Rodolfi et al.0 2023 b b b b a To be determined by EPA through a future rulemaking.65 0.0 2018 7.0 28. microalgae have demonstrated potential oil yields that are significantly higher than the yields of oilseed crops Many pathways are currently under consideration for (Exhibit 1.0 2021 13.95 2012 0. while algae may co-products within biorefineries offer comparable biomass productivity as lignocellulosic feedstocks – the key biomass resource factored in the study.75 18. CELLULOSIC BIOMASS-BASED ADVANCED BIOFUEL TOTAL RENEWABLE BIOFUEL DIESEL REQUIREMENT FUEL REQUIREMENT REQUIREMENT REQUIREMENT 2009 N/A 0.0 a 2. The most promising among these are routes from soybeans. b To be determined by EPA through a future rulemaking. Significant acreage and productivity will be required • Minimizes competition with conventional agriculture for biomass production to generate sufficient feedstock • Utilizes a wide variety of water sources to meet the RFS mandates.25 22. offer significant advantages that complement traditional feedstocks towards these fuels. Overview  .0 33.1 2010 0.5 a 15. approximately 15 times more productive to advanced biofuels such as high energy density fungible than jatropha.0 2020 10.1 0.5 2016 4.75 a 3.25 0. the study did not explore the potential of algae.

dates back to the 1940s.. a Adapted from Chisti (2007) b Estimated yields. The program lasted from 1978 until 1996 and supported research primarily at DOE’s National Exhibit 1. genetic Jatropha 202 engineering. The isolates were screened for their tolerance to variations Although. and also for their ability not yet emerged. this report The Aquatic Species Program researchers collected more than 3. (1978) indicated that algal • Algal productivity can offer high biomass yields per systems could produce methane gas at prices competitive acre of cultivation.2 Comparison of oil yields from biomass feedstocksa Renewable Energy Laboratory (NREL.S. 1. process development. The Aquatic Species OIL YIELD Program also funded research at many academic CROP (GALLONS/ACRE/YR) institutions through subcontracts.Advantages of Algal Feedstocks Early Work to 1996 Algae can be preferred feedstock for high energy density. The collection was narrowed approaches to the technologies being explored today. Proposals to use algae as a means of producing energy fungible liquid transportation fuels. Some of the highlights are described briefly below. Overview  3 . and temperature. northwestern. freshwater supplies. The concept of utilizing the lipid stores as a source of • Algae can recycle carbon from CO2-rich flue emissions energy. During the early years. Various reports during the 1950s • Algae can utilize waste water.000-6. a scalable. but the focus changed to Camelina 62 liquid fuels (biodiesel) in the early 1980s. thereby reducing competition for limited as nitrogen or silicon. with projected costs for fossil fuels. could lead to this phenomenon. the emphasis was on using algae to produce hydrogen. to the 300 most promising strains. primarily green algae (Chlorophyceae) and diatoms (Bacillariophyceae). Approximately $25 million (Sheehan.2 A History of Domestic and southeastern U. Advances were made through algal strain isolation and characterization. commercially viable system has in salinity. comprehensive research efforts to date on fuels from microalgae. and and 1960s indicated that starvation for key nutrients. 1998). There are several started in the late 1950s when Meier (1955) and Oswald aspects of algal biofuel production that have combined to and Golueke (1960) suggested the utilization of the capture the interest of researchers and entrepreneurs around carbohydrate fraction of algal cells for the production of the world: methane gas via anaerobic digestion. earlier studies have laid foundational to produce neutral lipids. Techno-economic analyses Oil palm 635 and resource assessments were also important aspects of the program. The discovery that • Algae cultivation strategies can minimize or avoid many species of microalgae can produce large amounts of competition with arable land and nutrients used for lipid as cellular oil droplets under certain growth conditions conventional agriculture. inland saline habitats that typically The advantages of algae as a feedstock for bioenergy undergo substantial swings in temperature and salinity. such saline water. pH. and demonstration- scale algal mass culture. gained serious attention only during from stationary sources. representing a diversity of aquatic Algal Biofuels Development environments and water types. 1998) was invested during the 18-year Soybean 48 program. A detailed engineering analysis by Benemann et al. have been apparent since the mid-twentieth century. • Algal biomass is compatible with the integrated biorefinery vision of producing a variety of fuels and The Aquatic Species Program represents one of the most valuable co-products. In 1998. a comprehensive overview Algae 1.000 strains of microalgae over a seven-year period from various sites in the western. 1. produced water. surges through the decade. including power plants and the oil embargo of the early 1970s and the energy price other industrial emitters. however. this idea ultimately became a major push of DOE’s Aquatic Species Program. Sunflower 102 studies of algal physiology and biochemistry.500b of the project was completed (Sheehan et al. Many of the strains were isolated from shallow. formerly the Solar Energy Research Institute).

In this species. For example. estimated costs for unextracted 1998). These initial to be more than 90% with careful pH control. 1987 and in desert regions of the Southwest (Maxwell et al. A Test Facility in Roswell. stable production of algal successful transformation of microalgae (Dunahay et al. purification. 1985). The termination of many billions of gallons of fuel. operated by key enzyme is acetyl-CoA carboxylase (ACCase). biosynthesis of the storage carbohydrate. but Among various techniques. ACCase activity was found to increase deep). The 4 1. scale-up.and capital-intensive. productivity improvements. Under Aquatic Species Program subcontracts. biomass was demonstrated. shallow (10-20 cm for TAG synthesis. chrysolaminarin saline water.. issues associated with such methods. was the focus demonstration-scale outdoor microalgal cultivation of many of the biochemical studies. Harvesting is a the single largest influence on fuel cost. The systems were characterized (Roessler.. but no further fuel much as 60% of their dry weight in the form of lipid.. as unlined open ponds (Benemann and Oswald.g. Inc.. it became possible to manipulate the expression of utilization (bubbled through the algae culture) was shown ACCase in an attempt to increase oil yields. 1999. In addition. some process options (e. petroleum consumption.. suggesting that the of the Aquatic Species Program in 1996 halted further technology could have the potential to have a significant development of these potentially promising paths to impact on U. 1988) with the hope of eventually turning down the Sufficient resources were identified for the production flow of carbon through these pathways. studying Program and shown to be one of the less challenging this response led to valuable insights into the mechanisms aspects of the technology. Conversion of Although nutrient deficiency actually reduces the algal oils to ethyl. 1989). the commercially viable strains for oil production.S. Cyclotella cryptica. the Roswell area. and conversion to fuel are critical to successful In addition. and CO2 resources (power plants). strains from the wild. smaller ponds for inocula production. and other needed for biological productivity and various problematic issues. growth was conducted in California. enzymes involved in the Aquatic Species Program. Cost-effective methods of harvesting the necessity for very low-cost culture systems. but lower oil producing. With the advent of the first successful in that long-term. Different cost process step that is highly energy. other biofuel of lipid biosynthesis.. plus several under the nutrient stress conditions (Roessler. further Aquatic Species Program focused on solvent systems.$186 per purification of the oil are also cost-intensive steps. Under inducing conditions. Studies focused on suitable land. 1990).. but studies examined the ability of many strains to induce failed to fully address the scale. However. and the efficiency of CO2 1995). primarily in C. but overall biomass productivity averaged no effect was seen on lipid production in these preliminary around 10 g/m2/day with occasional periods approaching experiments (Jarvis et al. such and dewatering algal biomass and lipid extraction. New Mexico. as biosynthesis of these compounds competes for fixed carbon units that might otherwise be used for Several resource assessments were conducted under the lipid formation.. or engine testing was carried out. production. One serious problem encountered was that the desired starting strain was often outgrown by faster Additional studies focused on storage carbohydrate reproducing. cost. The raceway design suggesting that it may play a role as a “spigot” controlling was based on the “high rate pond” system developed at lipid synthesis. 1988). and thus the enzyme was extensively University of California-Berkeley. an oleaginous diatom. distance from CO2 point sources. it became clear that novel solutions would be of aquifers. and environmental lipid accumulation under conditions of nutrient stress. and New Mexico under conditions of insufficient silicon (a component of (Sheehan et al. harvesting via flocculation even with optimistic assumptions about CO2 credits and was deemed particularly encouraging (Sheehan et al. which Microbial Products. analyses led to differing conclusions on fuel cost. cryptica were characterized (Roessler. conversion of lipids to gasoline) species in the collection were shown to accumulate as were evaluated (Milne et al. biological productivity was shown to have commercialization of the technology. 1990). costs of these resources can vary widely depending During the course of the Aquatic Species Program upon such factors as land leveling requirements. characterization. Sheehan et al. primarily triacylglycerides (TAGs) (Chisti. Extraction of oil droplets from the cells and algal oil were determined to range from $59 .. depth research. 50 g/m2/day. (Weissman et al. Low attempts at metabolic engineering identified a pathway to nighttime and winter temperatures limited productivity in modify the gene encoding in the ACCase enzyme.or methyl-esters (biodiesel) was overall rate of oil production in a culture (because of the successfully demonstrated in the Aquatic Species concomitant decrease in the cell growth rate). Detailed techno-economic analyses underlined process steps.000 m2 outdoor. 1998). Overview  . 2007). however. 1998). paddlewheel-mixed raceway ponds. Hawaii. This catalyzes the first step in the biosynthesis of fatty acids used facility utilized two 1. 1996).After promising microalgae were identified. Of particular note was the Outdoor the cell wall) is a trigger for increased oil production.

and several DOE Small Business Innovative to reduce the level of risk and uncertainty associated with Research (SBIR) request for proposals. Cyclotella cells. In particular.nrel. it also became clear that significant barriers would need to be overcome in order to achieve an economically feasible process. However. 1987). federal agencies.3 Algae-to-Biofuels: Program. researchers concluded that algal-derived Algae must be considered as part and parcel of the SNG would not be cost-competitive with fossil fuel gas. the Air Force Based on the information provided at the Workshop. and Biomass Program are all funding research activities By reviewing the technology gaps and cross-cutting needs. a number of U. the National to tackle the challenges of algal biofuels is thus Science Foundation. 1. Primary efforts were focused on open ocean culture of Abundant. and public-private partnerships. cyanobacteria. Similar to the findings of the Aquatic lifeblood of the burgeoning biofuels industry today. Office of Science. policymakers. these activities must be accompanied with DOE’s Advanced Research Projects Agency-Energy conducive developments on the non-technical fronts – (ARPA-E). 1998).gov/publications.barrel (Sheehan et al. funding levels are beginning to increase. significantly outpacing government funding. such as a major Defense Advanced Research Projects Agency solicitation. there is no parallel agricultural Since the end of DOE’s Aquatic Species Program in enterprise equivalent for cultivating algae at a similar 1996.S. and sustainable feedstocks are the California kelp. Office of Fossil Energy. In contrast to the development of cellulosic biofuels which benefit from a direct agricultural and process Research from 1996 to Present engineering lineage. Also from 1968-1990. that include investigating microalgae. It was concluded that algal biofuels would not be cost-competitive with petroleum. federal funding for algal research has come scale. needed to support commercialization activities. Additionally. A sizable and strategically structured investment from DOE. State funding programs and research support from private Technology and Analysis Challenges industry also make up a significant proportion of research funding. cost-effective solutions for the culture and process engineering challenges. and the private sector in implementing a nationally coordinated effort toward Additionally. Overview  5 . Courtesey of CCMP. and the Department of Agriculture.. it was Office of Scientific Research (AFOSR) algal bio-jet determined that a great deal of RD&D is still necessary program. the Aquatic Species Program was successful in demonstrating the feasibility of algal culture as a source of oil and resulted in important advances in the technology. Private investment in algal biofuels This Roadmap seeks to lay down the first comprehensive has been increasing at a dramatic rate over the last few state of technology summary for fuels and co-products years. from algal feedstocks and to document the feasibility and techno-economic challenges associated with commercial scaling up of processes. and the Roadmap aims to guide researchers and engineers. feedstock mix for producing advanced biofuels. a research initiative to determine the technical and economic feasibility of macroalgae cultivation and Opportunity and Challenges conversion to fuels. DOE sponsored the Marine Biomass 1. which was trading at less than $20/barrel in 1995. the Department of Defense. affordable. particularly to substitute natural gas Today (SNG) via anaerobic digestion (Bird and Benson. macroalgae for biofuels and beneficial re-use of CO2. suggest that the algae-to-biofuels process so it can be commercialized. Further. the work highlighted the need to understand and optimize the biological mechanisms of algal lipid accumulation and to find creative. national labs are developing a viable and sustainable algal biofuel industry. Detailed results from the Aquatic Species Program research investment are available to the public in more than 100 electronic documents on the NREL Web site at www. increasingly focusing on algal biofuels research. Recent initiatives. Overall. regulations and standards. Species Program.

. closed.. energy. direct fuel production. heterotrophic. and thermal management PURSUING STRATEGIC R&D: TECHNO-ECONOMIC MODELING AND ANALYSIS Given the multiple technology and system options and their interdependency. FEEDSTOCK and mixotrophic growth) Algal • Achieve robust and stable cultures at a commercial scale Cultivation • Optimize system for algal productivity of fuel precursors (e. 6 1. phototrophic. and separations CONVERSION CONVERSION • Investigate multiple approaches to liquid transportation fuels (e. solvent systems.g..g. carrying capacity. and materials from algal remnants (e. centrifugation. and surfactants) Co-products • Optimize co-product extraction and recovery • Conduct market analyses. animal/fish feeds. an integrated techno-economic modeling and analysis spanning the entire algae to biofuels supply chain is crucial in guiding research efforts along select pathways that offer the most opportunity to practically enable a viable and sustainable algae-based biofuels and co-products industry. and coastal/off-shore systems. lipids) • Sustainably and cost-effectively manage the use of land. biogas. and nutrients • Identify and address environmental risks and impacts • Investigate multiple harvesting approaches (e. and mechanized seaweed harvesting) Harvesting and • Minimize process energy intensity Dewatering • Lower capital and operating costs • Assess each technology option in terms of overall system compatibility and sustainability • Investigate multiple approaches (e. pressure. biofuel. intermediates.g. activity. OVERCOMING BARRIERS TO ALGAL BIOFUELS: TECHNOLOGY GOALS PROCESS R&D CHALLENGES STEP • Sample strains from a wide variety of environments for maximum diversity • Develop small-scale. fertilizers. bioplastics. sonication. filtration. side reactions. and end-product variability and Utilization • Optimize distribution for energy and costs in the context of facility siting • Comply with all regulatory and customer requirements for utilization (e. and secretion) Extraction and • Achieve high yield of desired intermediates. such as operational temperature. sedimentation. energy. biochemical conversion. Overview  .g. supercritical fluid... and durability Conversion • Reduce contaminants and reaction inhibitors • Minimize process energy intensity and emissions over the life cycle • Achieve high conversion rates under scale-up conditions • Identify and evaluate the co-production of value-added chemicals.. climate. selective extraction. flocculation. subcritical water. water & nutrient reuse.g. water. high-throughput screening technologies Algal Biology • Develop open-access database and collections of existing strains with detailed characterization • Investigate genetics and biochemical pathways for production of fuel precursors • Improve on strains for desired criteria by gene manipulation techniques or breeding FEEDSTOCK • Investigate multiple approaches (i. stability. dissolved air floatation. industrial enzymes. thermochemical/catalytic conversion. hybrid. and anaerobic digestion) Fuel • Improve catalyst specificity. open. water. microwave. engine performance and material compatibility) • Assess and characterize land.g. and nutrient resource requirements for siting of microalgae Resources (heterotrophic & photoautotrophic) and macroalgae production systems and Siting • Integrate with wastewater treatment and/or CO2 emitter industries (in the case of heterotrophic approach) • Address salt balance.e. including quality and safety trials to meet applicable standards • Characterize algal biomass. energy balance. and bioproducts under different storage and transport scenarios for INFRASTRUCTURE INFRASTRUCTURE Distribution contamination. weather impacts. preserve co-products Fractionation • Minimize process energy intensity • Investigate recycling mechanisms to minimize waste • Address scaling challenges..

A. P. 223–242. with algal biofuels commercialization.E.. ORNL/TM-2006/66. 31(6). Oak Ridge. & Golueke. regulation. Roessler.G. Jarvis. 1999. However. However. NTIS# H CP/ T. R. Engineering Design and Cost Analysis of a Large-Scale Microalgae Biomass System. Dunahay. and policy and regulatory challenges as that RD&D activities need to be carried out under a resolution of these issues will likely require participation framework of standards.J. Effects of silicon deficiency on lipid composition and metabolism in the diatom Cyclotella cryptica.. 2005. Journal of Biological Chemistry.. Available at: [Accessed November 3. & Ohlrogge.G. Isolated gene encoding an enzyme with UDP-glucose pyrophosphorylase and phosphoglucomutase activities from Cyclotella cryptica .. with shape-selective zeolites. 1993. E. P. Microalgae for oil: strain selection. T. C. E.J. 2008. Oak Ridge National Laboratory.. 1.. W.. & Nagle. J. P. Being a nascent industry. 1955.G.US Patent 5928932 Abstract. Roessler.. Solar Energy Research. R..Regulations and Standards Public-Private Partnerships While the Roadmap’s primary objective is to highlight A collaborative framework of public-private partnerships the technical challenges and opportunities associated offers an opportunity to jointly address the technological. 11. structuring public- biofuel developers need to foresee and understand the private partnerships for successful ventures is a challenge potentially applicable legal requirements early on in the in itself given the myriad issues and interests.G. & Roessler. Pursoff. such as research and development process to help ensure algae intellectual property rights. 1960. Biomass.e. J. R. Biological transformation of solar energy.L. Y. 219-232. Biotechnology Advances. References Benemann. Chisti. Overview  7 . Jarvis. Cloning and characterization of the gene that encodes acetyl-coenzyme A carboxylase in the alga Cyclotella cryptica. Final Report to the US Energy Department. Perlack. Biodiesel from microalgae. 394–400. Meier. 24. N. 1004–1011. 179–183. there are no existing standards for various aspects of algal biofuels production. Journal of Phycology. & Roessler. 25(3). biofuels and co-products) comply with applicable workforce needed to allow this industry to grow. Appl. 1995. 1988. induction of lipid synthesis and outdoor mass cultivation in a low-cost photobioreactor. 1605(UC-61).D. Biomass as feedstock for a bioenergy and bioproducts industry: the technical feasibility of a billion-ton annual supply. & Oswald. 2007. Evans. Biological cycles in the transformation of solar energy into useful fuels. it is recognized economic.. consumption standards.. et al. Biotechnol Bioeng. 102(1). Catalytic conversion of microalgae and vegetable oils to premium gasoline. 1990. W. Adv. Microbiol.. 100–112.G. P. Milne. 294–306. T. Oswald. Rodolfi.J. 91..R..B. P. Phycol. 21(3). Also. supporting education are legally and safely developed and the end-products will be critical to create intellectual talent and the (i. Algal from multiple entities.. and policy. 2009]. Genetic transformation of the diatoms Cyclotella cryptica and Navicula saprophila. 1978. TN. J. et al. L. RD&D activities can inform further development of applicable laws and standards..

and Algae can be isolated from a variety of natural aqueous nutrient requirements. distribution but also for the temporal succession brought about by seasonal variations of algae in their habitats. type and strain of algae being cultivated will ultimately However.. locations to maximize genetic diversity. products. (seaweeds). they typically live in aquatic environments and are capable of photosynthesis. 2005). which includes fresh.or eukaryotic. Geographic Information Systems (GIS) data and and carbohydrates.e. on the cultivation approaches and fuel precursor desired. Algae can be in suspended mass cultures. pH. 2005). Screening Criteria and Methods new strains should be isolated from a wide variety of An ideal screen would cover three major areas: growth environments to provide the largest range in metabolic physiology. and soil (Round. Due to their diverse characteristics.2. CO2 levels). oxygen levels. cultivation and strain development. actual strain identification should be based on The goals of algae isolation and screening efforts are to molecular methods such as rRNA sequence comparison. algae are organisms from different kingdoms of life. rivers. metabolite production. large-scale sampling efforts should be all parameters simultaneously would be helpful. The exact screens employed would depend geothermal springs. Because it is not yet known how algae will be cultivated on a mass scale. Some of these could prove to be valuable co- of ownership of isolated strains should be considered. In all cases. The selection of specific locations can be determined by sophisticated site Screening for metabolite production may involve selection criteria through the combined use of dynamic determining the cellular composition of proteins. versatility possible. lipids. this is not always the case. 2.1 Strain Isolation. ponds. Because all of these parameters habitats ranging from freshwater to brackish water. and cyanobacteria (historically known as traditional cultivation techniques may be used such as blue-green algae). Algal Biology The term “algae” commonly refers to a diverse mix of Additionally. some algal strains take weeks to months to affect every step of the algae to biofuels supply chain. the enrichment cultures (Andersen and Kawachi. and strain robustness. see Anderson. and measuring the productivity of analysis tools. 8 2. can include public lands as well as various sites within many strains also secrete metabolites into the growth our national and state park systems. whereas benthic algae may single or multi-cellular and pro. of automated systems that provide information regarding Furthermore. and though find application in biofilm-based production facilities. lakes. Natural Habitats maximum cell density. Because of Isolation and Characterization of morphological similarities when comparing many algal Naturally Occurring Algae species. brackish. Furthermore. a helpful screen for oil production would terrestrial environments in a variety of geographical allow for distinction between neutral and polar lipids. and generation. marine require significant experimental effort. Algal Biology  . Ecosystems to be sampled could include the organism regarding metabolites useful for biofuels aquatic (i. tolerance to environmental variables (temperature. maps. typically found in planktonic (free floating) and benthic algae have been unified based on their ability to carry out (attached) environments. within an aqueous habitat. For large-scale sampling and isolation efforts. 2004). coordinated to ensure broadest coverage of environments and to avoid duplication of efforts. The term growth physiology encompasses a number of parameters such as maximum specific growth rate. hypersaline. other gene markers. Planktonic algae may be used photosynthesis and live in aquatic habitats. Traditionally. and new approaches are needed to develop Sampling strategies should not only account for spatial screening methods for extracellular materials. the development and hyper-saline environments. or identify and maintain promising algal specimens for in the case of closely related strains. oceans. 1984). acidic and alkaline environments) and For example. salinity. be isolated by traditional methods (for a comprehensive review of algal culturing techniques. high-throughput automated isolation techniques involving Screening and Selection fluorescence-activated cell sorting (FACS) have proven to be extremely useful (Sieracki et al.. streams. Collection sites and would provide fatty acid profiles. Algae include Isolation Techniques microalgae (unicellular eukaryotic organisms). questions medium. macroalgae For isolation of new strains from natural habitats.

It could also reduce dissemination of detailed information about potentially the number of redundant screens of algal species. They currently maintain thousands & screening results of different algal strains and support the research and industrial community with their expertise in algae 5. algae stems from a lack of high-throughput methodologies because many of the strains in these collections have been that would allow simultaneous screening of multiple cultivated for several decades. Transcriptomics. pathogenicity. Role of Culture Collections as National reference) Algae Data Resource Centers 2. The function of a culture collection often 6. pH) & support research on determining strain characteristics. and provide 3. only a few major collector) algal collection centers exist in the United States and 4. no central database exists that provides global 9.. Omics data (Genomics. it is also industry has further exacerbated the flow of relevant strain important to consider the strain’s robustness. that can be used for mass culture in biofuels applications. native al. with technologies is an important step in enabling the testing about 3. Texas). Not only document data on strains. Some minimal growth information is available from includes parameters such as culture consistency. it has not yet been established if these variety of algae for assembly into a culture collection that methods are valid across a wide range of algal strains (Elsey will serve as a bioresource for further biofuels research. but it requires a significant quantity of biomass (Ahlgren et it would be prudent to consider the isolation of new. At present. and breeding and metabolic engineering approaches to improve though these methods require much less biomass (as little biofuels production. which data. such as growth rate and metabolite properties. The culture collections such as UTEX (The Culture Collection development of small-scale but high-throughput screening of Algae at the University of Texas at Austin. and CCMP (The Provasoli-Guillard of hundreds to thousands of different algal isolates. they could potentially are rapid screening procedures necessary for the biofuels serve as nuclei for the development of national algae field. Culture collections are necessary to preserve the diversity preservation) of natural habitats. To obtain versatile and robust strains common method for determination of lipid content in algae. molecular. basic research resources. it will be important to isolate a wide as a single cell). Algal Biology  9 . metabolites. Information could include: 1. Plasmids and Phages transcends simple depository functions. Strain properties: Cytological. Environment and strain history (specific habitat. community stability. Therefore. Further improvements in analytical methodology could be made through the As the major culture collections already collect and development of solid-state screening methods. Strain name (species. germination conditions cryopreservation. cultures (Sheehan et al. 1959). They may also 7. biochemical. protect genetic material. the bottleneck in screening large numbers of Harbor. some may have lost original phenotypes. other countries. de la Jara et al. Strain administration (number in collection.. Biological interaction (symbiosis. The establishment studies revealed that algae strains tested in the laboratory of a central strain. Solvent extraction. Proteomics. and susceptibility to more detailed information on growth.For mass culture of a given algal strain. existing culture collections. Growth conditions (media. ) (Bligh and Dyer. However. or in connection with outside collaborators. and phylogeny either by themselves 8.. with more than 2. temperature. Fluorescent methods strains directly from unique environments. and predators present in a given environment. For both direct using lipid-soluble dyes have also been described. for example. algal strains. Mutants biology. such as mating capability or versatility regarding productivity. but it is very difficult to obtain resilience. Previous robustness of particular existing strains. Protection of intellectual property in private or Metabolomics) 2. et al. but could prove extremely useful for the identification resource centers. 1991. small-scale simulations of mass A number of algal strains are currently available from culture conditions will need to be performed. National Center for Culture of Marine Phytoplankton at the Bigelow Laboratory for Ocean Sciences in West Boothbay At this time. Maine). valuable strains. taxonomy. subspecies name. 2003). toxicity) Currently. 2007. is the most nutrient requirements.500 strains. Culture collection organizations of species (particularly in mixed field samples) necessary could be responsible for the gathering and for the future of algal ecology. Practical applications (general and industrial) information on the characteristics of currently available 10.000 strains. open access repository could accelerate do not always perform similarly in outdoor mass R&D of algae-based biofuels production systems. to determine a strain’s robustness. 1998)..

food low. if the desired product is volatile.. although there are algae known processes since the basic tools are in place. The ability of an algal species to secrete fuel precursors may be attractive because it could reduce or skip the cell In relation to biofuels. Co-products could include pharmaceuticals manufacture everything from cancer drugs to high. but we cannot be certain that laboratory strains unfavorably alter fluidics properties or provide a carbon and production strains will be sufficiently related to allow source favoring growth of contaminants. Rapid growth is important both for overall productivity and the ability to compete with Capability for Heterotrophic or contaminating strains. 2006).cost commodities such as lysine and ethanol. this is not generally a problem with well- isoprenoids. 2006). so it is possible hydrocarbon phase in solution (Banerjee et al. and thus are not free to form an organic large-scale culturing (Sheehan. Pilot-scale experimentation and further metabolic engineering is required to evaluate Culture stability over long periods will be a key to low cost possible advantages and disadvantages of secretion. Botryococcus was shown in the Aquatic Species Program that not all braunii). A reasonable first approach to identify model species that are optimal for the production of a desired fuel is through a survey of the literature. For example. or materials for nanotechnology (in the case of the silica cell wall of diatoms). they are still associated with the cells in a lipid strains that grow well in the laboratory are suitable for biofilm matrix. an abundance of exported lipids could feasible. or a screen of environmental isolates for species 10 2. supplements.. 1998). Other traits like the ability to grow to Mixotrophic Growth high cell density in continuous culture may allow a strain to be maintained while at the same time reducing the amount Heterotrophic or mixotrophic growth capabilities may of water to be processed daily. secretion for lessons from the former to be applied to the latter. In such a strain. or co-product to be produced. alcohols (either directly or established fully-heterotrophic fermentation technologies through biomass conversion). in a practical sense. the number of algal systems that can be studied in depth has to be limited because a critical mass of researchers Secretion of Products or Intermediates is required to make progress on a given species. lipids. However. systems could be studied. intermediate. Even that other strains will be chosen for production. source. production of biofuels. However. even under mixotrophic agents could reduce the costs of harvest as long as it could conditions where the substrate is not known to be be controlled to avoid settling in the cultivation system. to providing information on basic cellular processes then collection of the atmosphere above the culture will regarding the synthesis of fuel precursors. there are two types of algal model harvesting step. Species with sequenced genomes and transgenic secretion actually makes the product more readily capabilities are the most amenable to investigating cellular available. However. Algal Biology  . However. contamination by undesired microbes living off the carbon Possible fuel types of interest could include H2. there may be practical problems systems to consider studying: species or strains amenable to consider.. If the carbon source is utilized by the cell. a large number of model and possible strain development for production. Adapting if sustainable secretion could be achieved. cellular metabolism is already geared toward production.Selecting Algal Model Systems that naturally make abundant amounts of the desired for Study product. or methane (via anaerobic that are currently deployed worldwide at massive scale to digestion). growth in both Targeting Desired Fuel Product or Intermediate light and dark periods is possible. Also the ability addition of supplemental carbon results in increased lipid to flocculate without addition of chemical flocculating accumulation (Xu et al. carbohydrates. and high cell densities can be achieved. viruses could also be a useful phenotype. such as. For example. and species be necessary to isolate it. Finally. Resistance to predators and be attractive attributes of algal strains. which simplifies characterization Given the diversity of algae. of either intermediates or products into the growth medium could make these compounds vulnerable to contaminating Useful Algal Characteristics microbes for catabolism. it is not clear the lessons learned from laboratory model strains to what the effect of a lipid emulsion in an algal culture would strains known to be capable of large-scale growth may be be. In some species. transported into the cell (Ceron Garcia et al.g. which will necessitate the use or strains with characteristics useful for large-scale of closed bioreactors. 2002). A potential disadvantage of the addition One consideration in choosing model systems is the of external carbon sources is the possibility of increased type of fuel. it to secrete long-chain hydrocarbons (e. Also to be considered is whether growth.volume/ (therapeutic proteins and secondary metabolites).

important to strain development and designing cultivation strategies. a link between starch and lipid metabolism has There are many good reviews available that cover basic been established. 2002). Ruuska tend to absorb all the incoming light. therefore. It may be useful to carry out photosynthetic of carbon between alternative storage products will efficiency studies in algae similar to those that have be important for metabolic engineering of algae. in their productivity. cells nearer to the light source accumulation phase (Kang and Rawsthorne.. Cyanobacteria and macroalgae. understand the metabolic pathways and processes that Understanding carbon partitioning will require extensive generate them in order to advance biofuels production. exposed to high light. 2004.. pathway analysis. biomass 2007. 2002). carbohydrates. Under stress conditions such as high light or (Long et al. Yang et al. the percent of biomass and Fernie. it was shown that such as triacylglycerols (TAG) as their main carbon storage reducing the size of the chlorophyll antenna can increase compounds.. Certain microalgal species also naturally the efficiency of light utilization (Polle et al. A large majority of and Biochemistry absorbed incident light is dissipated as heat and could be Many algae are photosynthetic organisms capable of considered “wasted. for biofuel production. algal photosystems have built-in More recently. 2007). a potentially important implication limitations of algal photosystems regarding light utilization.e.. A fundamental understanding of both cultivation process design and strain improvement ‘global’ regulatory networks that control the partitioning projects. Though theoretical However. with lipid these processes if we are to better engineer the capture accumulation in macroalgae typically being less than 5% of and utilization of light energy for biomass production. 1994.. carbon partitioning in algae is less understood biomass productivity values in the range of 100-200 g/ and research on how algal cells control the flux and m2/day have been presented (Chisti. preventing it from et al. Allen et al. indicating that starch is an important storage reaching more distant cells (Christi.. nutrient starvation. Mcdermid and Stuercke. therefore. 1994. 2008). possible that TAG and starch could to maximize light exposure (see Chapter 3). and predictive Photosynthesis and Light Utilization modeling (Vemuri and Aristidou. and shares the Eberhard et al. which can lead to oxidative damage. are all potential biofuels lipids and/or carbohydrates could be very useful for biofuels or biofuel precursors. Metabolic networks have been reconstructed in various microbes from genomic and transcriptomic data. Foyer et al. there is no partitioning of photosynthetically fixed carbon into various current consensus on the true maximum productivity of groups of major macromolecules (i. the been done in plant systems to understand carbon flux in efficiency of photosynthesis is a crucial determinate biosynthetic and degradative pathways (Lytovchenko et al. plants (Hu et al. Libourel and Shachar-Hill. algae. as 2008). proteins.. Lipids and carbohydrates.. Nelson and Yocum. It is. studies in higher plants showed that when 2. Schwender et al. and Niyogi. and potentially. Algal Biology  11 . More emphasis should be placed on understanding a general rule. In and starch metabolism was most active before the lipid high cell density cultures. Starch is a common carbon and energy algal photosynthetic processes (Nelson et al. Theoretical productivity is an important concept. total dry weight (Mcdermid and Stuercke. Regardless of the cultivation practices used 2. accumulate large amounts of TAG (30-60% of dry weight). that is the desired fuel precursor. accumulate mostly carbohydrates. Research has also When algae are cultivated photosynthetically.. Krause and same precursors with the storage lipid TAG (Exhibit Weis. 2007 and 2008) Further.2. 1994. 1991).. storage compound in plants and algae. 1994. In young Arabidopsis seeds The majority of light that falls on a photosynthetic algal and Brassica embryos. affecting growth rate. 2008.. knowledge of metabolic pathways. 2005). 2002). 2007). photoinhibition or the reduction O2 and organic macromolecules such as carbohydrates of photosynthesis due to light damage can still occur and lipids. however. 2003). along with biologically Knowing how and when carbon is partitioned in a cell into produced hydrogen and alcohols. Even when compound and its synthesis precedes oil accumulation. 2009. 1999)... although concentrations approaching 20% lipid have been reported Carbon Partitioning and Metabolism in some species (Chu et al. because it can be used to set achievable goals for 2009. 2003. starch transiently accumulated culture at greater than laboratory scale is not utilized. and exhibit photosynthetic efficiency and lipid production There is still much to learn about the dynamics and at least an order of magnitude greater than terrestrial crop regulation of the photosynthetic apparatus (Eberhard et al.. It is. some microalgae accumulate lipids In an effort to overcome this barrier. been carried out for plants (Zhu et al. and lipids) is critically needed (Boyle and Morgan. 2008). Sweetlove production.2 Algal Physiology strategies to prevent the over-absorption of light energy. 2005. under harvesting solar energy and converting CO2 and water to certain light regimes.” Even with this photoprotection.1). 2003). 2006. there remains be inter-convertible..

TAG is formed by food reserves. and and interaction with lipid metabolism in algae. There are also many algae that completely lack 2000. mannitol. Some algae. or efficiently extracting biofuel precursors of TAG (Hu et al. These structures can be an important source of that is unique to TAG biosynthesis (Lung and Weselake. feedstock potential. though their physical structure and the presence in an acyl CoA-independent way (Arabolaza et al. degradation. must typically 2006. be fruitful to Most cyanobacteria have a peptidoglycan layer and cell further research de novo starch synthesis. are 3-phosphate. transferred from CoA to positions 1 and 2 of glycerol- for example alginate found in many brown algae. the nature of the interaction is unknown. biochemical feedstocks for conversion to liquid fuels.. 2002). Diatoms are also unique among algae for the presence of silica in their cell walls. plants and yeast based. envelope similar to those of gram-negative bacteria. 2000). therefore. some red algae and dinoflagellates. In the final step of TAG synthesis. 12 2.. are encased in a polysaccharide sheath (Hoiczyk and Hansel. Cell walls can also be a technical barrier. Another important consideration in algal strains is the a third fatty acid is transferred to the vacant position 3 composition and structure of the polysaccharide cell of DAG by diacylglycerol acyltransferase. as example many brown algae and diatoms. accumulate significant quantities of neutral storage lipids Many green microalgae are plant-like. and thus may play an for example. these phosphatase releases diacylglycerol (DAG). it is important to Primary Pathway for TAG Synthesis determine the predominate types that are present. 2008. greatly. featuring rigid such as triacyglycerols (TAG). (Roessler. including de novo fatty acid synthesis in the stroma of plastids. plant embryos or absence of other structural polysaccharides varies or seeds accumulated 40% less oil (Periappuram et al. many membrane lipids and/or carbohydrates to TAG have algal cell walls from different groupings are cellulose. As mentioned above. accumulate well as of neutral storage lipids such as TAG. when trying to access DNA for genetic important role in determining the final acyl composition manipulations. 2004). Yoo et al. Dephosphorylation of PA catalyzed by a specific end product (Bird and Benson. or fucoidin as 18 carbon fatty acids as precursors. while some algae lack synthesis. the redox balance favors formation of pyruvate as the 1995).. The acyltransferases be broken down into simpler sugars before conversion involved in TAG synthesis may exhibit preferences into biofuels.. naturally or under stress conditions. via three sequential acyl transfers (from acyl CoA) These major storage polysaccharides represent potential in the endoplasmic reticulum (ER) (Exhibit 2. While these results provide cellulose and have other polymers that provide structure an indication that starch synthesis is linked to lipid to the cell (Raven et al. 1971. and the Algal Carbohydrates technical challenges these structures may present in strain Algae are incredibly diverse in the kind of simple manipulation.1).. been identified and partially characterized (Horn et al. Vigeolas et al. 1992). growing cultures. The major pathway for the their main carbohydrate storage compound. 2006). for example for fermentation to produce alcohols. Fatty acids produced in the chloroplast are sequentially 2000a and 2000b). an enzyme wall. Athenstaedt and Daum. it may be of interest to research these TAG intermediates. 1988). Several formation of TAG in plants first involves algae commonly use starch for energy storage. Cyanobacteria often store large quantities incorporation of the fatty acid into a glycerol backbone of glycogen (Chao and Bowen.. Microorganisms capable of fermenting laminarin and TAG biosynthesis in algae has been proposed to occur mannitol from Laminaria hyperborea to ethanol have via the above Kennedy pathway described in plants. which are important cellulose-based cell walls and accumulating starch as potential fuel precursors. use 16 or carbohydrates such as laminaran. for The synthesis of cellular and organelle membranes. but like those from plants. 2008). An important lesson is the recognition of the diversity of algal polysaccharides and cell walls. carbohydrates. recently been demonstrated in bacteria.starch synthesis was impaired or inhibited. and extraction processes. It could. resulting in the formation of the central considered less suitable for ethanol fermentation because metabolite phosphatidic acid (PA) (Ohlrogge and Browse. Some red In microalgae. and complex carbohydrates that they use for carbon storage and cell structure. Since polysaccharides may still prove useful as intermediates diglycerides are usually present in high amounts in rapidly to other types of conversion processes and final fuels. Other abundant polysaccharides. cell walls entirely. However. Other algae. 1987).. Alternative pathways to convert from cells in mass culture. for specific acyl CoA molecules. Algal Biology  . such an interaction has been also algae also have a thick extracellular matrix composed indicated by studies on the diatom Cyclotella cryptica of important products such as agar or carrageenan. If carbohydrates Lipid Synthesis and Regulation are to be used as fuel precursors.

Algal Biology  13 . involved in lipid metabolism in algae. As an example. Glc6P: Phospholipids glucose-6-phosphate. PDH: pyruvate dehydrogenase (putative pathways were proposed in dashed lines).. 2004). KAS: 3-ketoacyl-ACP.. Moreover.C18 CoA Figure 2. and other 2. PA and be true. Because fatty acids are common precursors for the Alternative Pathways to Storage Lipids synthesis of both membrane lipids and TAG. This has revealed that algal lipid metabolism may be different lack of understanding may contribute to why the lipid from that in plants. ER AGPPase: ADP glucose pyrophosphorylase. If the or culture conditions. it could be a challenge to extrapolate information DAG can also be used directly as substrates for synthesis learned about lipid biosynthesis and regulation in laboratory of polar lipids. enzymes and new scenarios for lipid production and strain improvement. pathways. algal cell coordinates the distribution of the precursors to and the relative contribution of these individual pathways distinct destinations or how the inter-conversion between to overall TAG formation may depend on environmental the two types of lipids occurs needs to be elucidated. 2008. 1998). DAGT PDAT? PDAT: Phospholipids: DAG acyltransferase. If this proves to have not yet been studied in algae. how the Algae may possess multiple pathways for TAG synthesis. the de-novo Kennedy Pathway. as indicated by the presence and/or yields obtained from algal mass culture efforts fall absence of certain pathways and by the size of the gene short of the high values (50 to 60%) observed in the families that relate to various activities (Riekhof et al. Accase: acetyl GDAT? CoA carboxylase. Stahl et al. such as phosphatidylcholine (PC) and strains to production strains. to develop hypotheses for strain improvement in algae. laboratory (Hu et al. Diacylglycerol (DAG) ER: Endoplasmic reticulum. 2000. CO 2 Glc6P Starch AGPPase Central Starch Glc6P 3-PGA 3-PGA Amylases Metabolic Pyruvate Mitochondria PDH CO 2 Pyruvate Oxaloacetate Acetyl -CoA Chloroplast ACCase TCA cycle Acetyl-CoA KAS I KAS II Glycolipids SAD Galactolipase? C16 . the annotation of genes involved in lipid The regulation of the synthesis of fatty acids and metabolism in the green alga Chlamydomonas reinhardtii TAG in algae is relatively poorly understood.. Analyzing different algae could help ability to control the fate of fatty acids varies among algal to elucidate the possible pathways of TAG synthesis: the taxonomic groups or even between isolates or strains.. de novo fatty acid and lipid synthesis should Understanding lipid regulation can help to maximize be studied in order to identify key genes. Similarly. Such pathways property of individual species or strains. ACP: acyl carrier protein. These pathways are worth investigating to use information regarding lipid biosynthesis in plants when developing strains for improved lipid production. Thus. 2005). Sheehan et al. if any. it will be difficult galactolipids. the potential pathway for lipid basal lipid and TAG content may represent an intrinsic formation from starch reserves mentioned earlier.1 Major pathways for fatty acid Phosphatidic acid and TAG synthesis in plants and algae 3-PGA: 3-phosphoglycerate. GDAT: putative Acetyl- glycolipids: DAG acyltransferase.. Triacylglycerol Lipid Body Dahlqvist et al.

as an electron sink under photo-oxidative stress (discussed 2002). to the obvious physiological role of TAG as a carbon and 2009). the default pathway in some mitochondria and glyoxysomes/peroxisomes. Because of the potential importance of TAG play an important role for cell survival under stress. Organelle Interactions and storage lipid formation warrants further study. The study the plastid where they are assimilated. transport chain under high light or other stress conditions. and et al. some algal cells stop division and accumulate TAG as the main carbon storage compound. and their interactions with other organelles excess electrons.lipids (Alvarez and Steinbuchel. The molecules their lipid composition dominates extracts obtained from (e. In addition to load or discharge their lipid cargo (Zehmer et al. as rapid lipid energy storage compound. Huang. thereby serving as a mechanism to been proposed that these alternative pathways that convert “detoxify” these compounds and deposit them in the starch. 1993. their apparently widespread envelopes. with acyl earlier). As mentioned Lipid Body Formation and Relationship earlier. 1998. Such work is also important in algae to distribution and their great abundance in many lipid- better understand the interaction among organelles as storing seeds. 1998). The thylakoids of chloroplasts are with secondary carotenoid synthesis in algae (Rabbani the main intracellular membranes of eukaryotic algae. Fatty acids can then Despite the economic importance of algae as a source of be either assembled into glycerolipids at chloroplast a wide range of lipophilic products. In order to better understand lipid to membrane lipids. phatidylethanolamine or stress conditions as degradation of chloroplasts occurs. Chloroplast membranes control the exchange of metabolites between the plastid and the cytoplasm. which direct physical continuities between lipid bodies and may in turn cause inhibition of photosynthesis and damage bilayer membranes. TAG synthesis may also phosphatidlyglycerol ( ~15%) (Harwood. Synthesis of Lipid bodies may dock with different regions of the ER TAG and deposition of TAG into cytosolic lipid bodies and plasma membrane.g. and spatial. sulfoquinovosyldiacylglycerol (~15%) and the chloroplast under stress. there have been relatively few studies on lipid glycerolipids assembled at the ER are then returned to bodies in algae compared with plants and fungi. with exceptions. Algal chloroplasts the carotenoid pathway are sequestered into cytosolic contain monogalactosyldiacylglycerol as their main lipid lipid bodies.. there are now doubts about it relates to lipid formation and lipid trafficking. toxic fatty acids excluded from the membrane system the fate of these abundant lipids remains unclear. excess membrane lipids. proteins.. including vitamins. and other components into form of TAG. This relationship may be both metabolic. Under utilize phosphatidylcholine. a close relationship is often may also play a more active and diverse role in the stress found between neutral lipids like TAG and the membrane response. Zhekisheva et al. Some acids.and glyco. Lipid trafficking of lipid-body biogenesis in plants has focused largely on is. with smaller amounts of digalactosyldiacylglycerol ‘sunscreen’ to prevent or reduce excess light from striking ( ~20%). 2002). It has as acyl donors. or astaxanthin) produced in cells under favorable growth conditions. In oil-producing microorganisms. β-carotene... 1992). the TAG synthesis pathway body accumulation occurs. and thus relax the over-reduced electron related to storage lipid formation requires further study. 14 2. 2008). the chloroplast stroma is the primary location to Other Organelles for fatty acid biosynthesis in plants. membranes or they can be exported to the ER and hydrocarbons and very long-chain ω-3 and ω -6 fatty assembled into lipids for cellular membranes. with growing evidence of induce over-production of reactive oxygen species. an important aspect of membrane formation the role of oleosins (Murphy. the role of oleosins in the biogenesis of plant lipid bodies. Nevertheless. metabolism in algae. therefore. lutein. Under high light stress. 2001). or with other organelles such as may be. The de novo TAG synthesis pathway can serve phospho. the formation of fatty acids could help consume bodies. It has been suggested that oleosins may be primarily Oxidative Stress and Storage Lipids associated with the stabilization of storage lipid bodies during the severe hydrodynamic stresses of dehydration Under environmental stress conditions (such as nutrient and rehydration that occurs in many seeds (Murphy. the exact relationship between oxidative stress. 2008).. starvation). the structure and function of lipid However. stress conditions on lipid production in algae. Carotenoid-rich lipid bodies serve as a (~50%). Algal Biology  .potential pathways to convert membrane phospholipids and The TAG synthesis pathway is also often coordinated glycolipids into TAG. and other macromolecules. excess electrons that and glycerol moieties exchanged between the different accumulate in the photosynthetic electron transport chain lipid classes. Current work in plants is understandable in view of their exclusive localization is focused on deciphering lipid transport across plastid on lipid-body surfaces. This and lipid fate (Benning. in order algae under stress conditions (Hu et al. cell division.

and proteomic studies. Colorado School of Mines ) 2. but the electron acceptor is switched at the level of ferredoxin (FD) from CO2 to protons under conditions that lead to H2 production. Three et al. and (d) inefficiencies in Biohydrogen the utilization of solar light energy (Seibert et al. cDNA microarray both nickel and iron (which are generally H2-uptake analysis.4 PC Chl fixation P700 0. Hydrogenases are the enzymes responsible for releasing molecular H2 (Ghirardi et al. Glycolysis ADP ATP starch pyruvate Acetyl-CoA + CO2 NAD++H+ NADH PFOR NPQR NAD++H+ FD(ox) FD (red) -1. fermentative H2 pathway coupled to starch production (Mathews and Wang. in the following reactions: 2H2O hydrogenases with improved tolerance to O2 (Cohen + light energy → O2 + 4H+ + 4e. As a substrate. (b) identifying metabolic pathways that pathways have been described in green algae: two light.2) (Melis et al. Posewitz. One of the most important characteristics also help provide information about photosynthetic of hydrogenases is that they are O2 sensitive. (c) engineering degradation (see Exhibit 2.. In all pathways.. studies involving Expressed containing iron (which are generally H2-evolving). metabolic Four biological challenges limiting biohydrogen production engineering through genetic manipulation represents in algae have been identified: (a) the O2 sensitivity of yet another strategy for the production of algal oils. compete with hydrogenases for photosynthetic reductant. hydrogenases.→ O2 + 2H2. Based on such information. 2005).Besides biochemical analysis to study algal lipids 2007). a These challenges could be addressed by (a) engineering potential fuel product. light.2 P680 Two are driven by light and the third occurs in the dark. Carbon is fixed under Photosynthesis normal photosynthesis with water as the donor. the light-driven pathways photosynthetic-electron-transport-coupled ATP production can either employ water (through photosystems II and (ATP is not required for H2 production). Algal Biology  15 .0 PQ Cyt b CO2 Cyt f 0. efficiency of solar light utilization (Polle et al. for example. 2000. and engineering their down-regulation during H2 independent. those and carbohydrates.2 Three different pathways for H2 production 1. ferredoxin (FD) is the primary electron donor to the hydrogenase enzyme. (c) regulatory issues associated with the over production of ATP. Either O2+2H+ water or starch can be the electron donor. carbon partitioning and lipid/carbohydrate synthesis in algae.8 Fx H2 Photosystem I P680* FA/FB FD H2ase Energy (Volts) -0. 1984). Some microalgae and cyanobacteria can produce H2. and a third.4 Pheo QA QB 2H+ Photosystem II 0. Gfeller the photosynthetic membrane for decreased efficiency of and Gibbs.2 P700* A0 A1 -0.8 Chl 2H2O Figure 2. 2003). There are two major types of hydrogenases. and (d) engineering I) or NADH from the glycolytic breakdown of stored the photosynthetic antenna pigment content for increased carbohydrate (through photosystem I). driven H2-photoproduction pathways. 2008). (b) competition for photosynthetic reductant at the level of ferredoxin.... would enzymes). M. or Sequence Tag (EST) analysis. 2009). (Drawing courtesy of Prof.

hexose) (Turner et al. While a better understanding of the basic for H2 production at night. 2002. genetic toolbox that can be used to modify process- 1987). and (c) learn to express multiple hydrogenases The generation and characterization of mutants is a so that electrons from both ferredoxin (Fd) and NAD(P) powerful approach to understand gene function and H can serve as electron donors to support H2 production.Recently there has been a focus on using cyanobacteria to 2. 16 2. al. potentially generate strains with desirable characteristics.. 1998). genetic and cellular processes involved in the synthesis another option for H2 production is to take advantage and regulation of potential fuel precursors from diverse of the fermentation pathways that exist in some algae species of algae. engineered than eukaryotic algae and have more O2. more knowledge cannot be overlooked. In all cases. For all of these with ethanol. All cyanobacteria examined thus far employ the relevant strains remains a significant technical hurdle. 1987). Methods to cultivate and process algae in photosynthesis has been demonstrated in the unicellular commercial settings are no less important to biotechnology. In cyanobacteria. and 3) highlights a few via nitrogenase or hydrogenase (Stal and Moezelaar. and the production of includes developing biohybrid (those with biological chemicals.. Genetic approaches are commonly also excretes typical fermentation byproducts including used to introduce genes. for commonly used bacteria. the limited algal to changing environmental conditions (Stal and Krumbein. From here. using the carbon reserves biology of algal growth and metabolite accumulation produced during the day. 2009). methods to manipulate of photosynthesis. Some of these methods can also be used to study the It is well established that dark fermentation suffers from localization of gene products (mRNAs and proteins) low H2 molar yield (less than 4 moles of H2 per mol within cells. modify genes or gene expression in a particular organism. fermentation is using modern analytical approaches will provide a wealth constitutive. the importance of being able to harness and synthetic components) and synthetic photosynthetic biotechnology approaches to generate algae with desirable systems that mimic the fuel-producing processes of properties for the production of biofuels and bioproducts photosynthetic organisms. the hydrogenase from is a potentially important element in the research effort Cyanothece PCC 7822 produces H2 in the dark and toward algal biofuels. Collectively. using nitrogenase as the catalyst. This is due to the production the toolbox allows traits to be recombined into a single of organic waste by-products described above along individual by mating parental strains. and CO2 (van der Oost et al. biological process engineering breakthroughs directly tolerant hydrogenases (Ghirardi et al. Similarly economic viability. ATCC 51142 (Toepel et and these are the subjects of Chapters 3 and 4. In order to fully realize the potential of H2 approaches. 2) describes were found to use pyruvate-ferredoxin oxidoreductase. 2008) and Oscillatoria (Stal and Krumbein. human health. Efforts should be undertaken to understand the fundamental To circumvent the inhibition of hydrogenase by O2.. examples of how algal biotechnology has been applied 1997). This temporal separation of H2 production from to date. the stability of the desirable trait through many production via indirect biophotolysis. this subsection seeks to 1) address the genetic of glucose to pyruvate. the ability to improve on native strains under non-N2 fixing condition.. While many of the challenges described above The biotechnology industry grew from more than 100 exist in these organisms. 1989). to delete or disrupt genes. questions to consider in the context of mass production. enabling technologies and analyses that can be applied which reduces ferredoxin for subsequent H2 production for biofuels and bioproducts. they are typically more easily years of basic biology and genetics R&D. 2008). and algae genetically remain far behind those developed hydrogenase structure and function is required.. Embden-Meyerhof-Parnas (EMP) pathway for degradation Thus. 2005). (b) remove or down-regulate competing fermentative pathways thus directing more of the cellular flux toward H2 Mutagenesis production. several cyanobacteria tools available to modify algal strains. Prince and Kheshgi. fungi and higher plants. Algal Biology  . and to acetate. cyanobacteria Cyanothece sp.3 Algal Biotechnology produce H2 (Tamagnini et al. However. hydrogen evolution pathways. accounting for their ability to adapt quickly of hypotheses for strain improvements. the unicellular non-N2-fixing cyanobacterium The Genetic Toolbox Gloeocapsa alpicola can evolve H2 from the fermentation Because biological productivity is a key driver for of stored glycogen (Serebryakova et al. several challenges generations and the possibility of unintended horizontal must be addressed: (a) improve photosynthetic efficiency gene transfer to other organisms are important research to increase the yield of carbohydrate accumulation. A possibility enabled new multi-billion dollar commercial enterprises to improve the efficiency of biological H2 production for agriculture. formate. Using hydrogenase as the catalyst. For algae that undergo sexual reproduction. Thus.

.. including resistance to neomycin. The green algae (Chlamydomonas. 1985. diatoms (Cyclotella. in the presence of an antibiotic). other approach is limited by the low frequency of naturally methods may need to be considered for production scale. which could exacerbate current problems developed. the decision as to which antibiotic standard chemical or UV-based mutagenesis approaches.. Navicula. Dunaliella. 1994). dinoflagellates (Amphidinium. Arthrospira). Chlorella. 1985. Any mutagenesis approach requires an appropriate screening technique to enrich for and isolate mutants. Anabaena.. 1995). 1995). For the latter. The mechanism of antibiotic native gene is to be entirely replaced). changes in fluorescence properties. A direct comparison of the by introducing a selectable marker randomly into two has shown that the nourseothricin system generates the genome (Adams et al. Nostoc. Mutants are more readily generated by For research purposes. and to select for cells in which element that functions in the species of interest. For selection could be used to generate useful algal strains complementation approaches. use of transposons (Miller and Kirk. 1989. Debuchy et al.. also been successful (Del Pozo et al. Dunahay et al. including cyanobacteria (Synechococcus. important consideration is that the use of antibiotics in Synechocystis.. generated by conserved ribosomal protein mutations have e. this resistance is a powerful tool for research. or introduction of resistance can also be an important factor. occurring mutations. appropriate mutations must without the need to generate genetically engineered be generated in the species of interest.. or through the larger numbers of transformants (Poulsen et al. antibiotic markers have been developed for microalgae. 2006). selection marker to use includes whether the antibiotic Drawbacks of this approach include the introduction compound is sensitive to light and whether its potency is of multiple mutations in a genome and the difficulty modulated by the salinity of the growth medium. kanamycin (Hasnain Targeted or tagged mutagenesis offers the advantage of et al. 2006). concern is the environmental implications of widespread Thalassiosira). 2005). presumably because requirements for expression levels of the gene are lower and less taxing to the cells. 1989). ideally in well- (GE) algae—something that could be desirable for characterized genes that can be easily complemented.. An algal strains.. large-scale algal production. simplified identification of the mutated gene. For example. 2002). a modified copy of the gene that inserts elsewhere into zeocin resistance requires stoichiometric binding of the the genome. 1993. Marker systems that take advantage of the ability to genetically complement Transformation Methods auxotrophic and metabolism mutants have also been Gene transfer systems have been established in many achieved (Kindle et al.... or that the native gene may need to Given a well-developed screening approach. Once an appropriate antibiotic resistance or Selectable Markers complementing gene is identified. which necessitates a large amount of screening.. whereas nourseothricin in gene expression.. constructs must be A powerful way to manipulate genomes is the ability to made to place the gene under control of an expression introduce DNA into the cell.. are examples of control elements that work across along with the DNA of interest on an extra-chromosomal evolutionarily diverse species (Dunahay et al.g. Tagging can be accomplished is inactivated enzymatically. 1999).g. Algal Biology  17 . However. This Sophisticated metabolic engineering could require the can include either a requirement for mutants to grow under introduction of multiple selectable or complementary certain conditions (e. element called a plasmid. Phaeodactylum. zeocin (Apt et al. but markers based on resistance assayed. large-scale production has two major drawbacks. Certain strategies can also enable changes antibiotic by the resistance protein. Dunahay et al.. This the DNA is present. Nelson et or increased neutral lipid accumulation via Nile Red al. spontaneous mutants arising from errors with increased antibiotic-resistant microbes. or markers. however. reduced chlorophyll fluorescence (Polle et al. The second Volvox). Most of the current selectable markers are derived to exhibit a characteristic phenotypic change that is easily from bacterial genes. 1999).. Hallmann and Rappel. first concern is the cost of the antibiotic. 1994). Typically. iterative be replaced in order to generate the phenotypic effect. Several in mapping the locus responsible for the phenotype. this is accomplished by typically involves using control elements from a introducing an antibiotic resistance gene as a selectable highly expressed gene in that species. 2.As long as an appropriate screening process is antibiotic use. to be expressed at a higher level than the native gene (Nelson et al. 1987) can be screening criteria. Antibiotic in DNA replication can be identified. However. and nourseothricin approaches rely on homologous recombination (if the (Poulsen et al. 1995).. Targeted 1996. there marker (Hasnain et al. Caveats are that the mutated gene may need staining (Cooksey et al.

but considering the is the biolistic (“gene gun”) approach (Armaleo et al.e. other inducible or repressible control elements would be useful for both research and commercial applications. selecting for functional RNAi can be problematic of microalgal cells generally integrates randomly into (Fuhrmann et al. sexual crossing are not well developed in microalgae. Algal strains often contain expression. Homologous recombination-based gene integration although there is a recent report of the method being approaches are common in many strains of cyanobacteria.. 2009).. 1990). Two general types of RNAi vectors can be constructed but this methodology could prove to be extremely – one containing an inverted repeat sequence from the important. only a small via homologous recombination is more desirable than percentage of selected transformants will have functional random integration because it can overcome phenotypic RNAi. which is useful for both nuclear and chloroplast slightly elevated.. intermediate. especially in the study of polyploid organisms multiple copies of their genome. use of single stranded DNA (Zorin et al. 1998). challenge to introducing DNA into a cell is the nature of allowing for precise control over the timing of gene the cell wall. microarrays). and another in which bidirectional only for a limited number of generations and must be transcription generates the double stranded RNA. high-throughput phenotypic screen for functional RNAi Successful recombination approaches have included the by co-targeting an amino acid synthesis pathway along addition of long flanking regions to the gene of interest with the desired gene of interest (Rohr et al. transgenes are overexpressed by A common method for introducing DNA into algal cells using strong control elements. Some diatoms can be propagated vegetatively gene to be silenced.. For most of these approaches. In a practical but less so in microalgae. 2005). The selection process was based on a gene is present. Many through double-stranded RNAs that are cut down to macroalgae species are capable of sexual reproduction. RNAi can inhibit transcription to improve commercial varieties of seaweed since the (Storz et al. 1996).Symbiodinium). Gracilaria. With the exception of cleaving specific mRNAs or sequestering them away Chlamydomonas.. ammonium (Poulsen and Kroger. The nitrate reductase promoter has proven cell wall. used in a red alga (Ohnuma et al. crossed periodically to maintain culture viability. RNAi approaches have been investigated primarily Homologous Recombination in the model green alga. Undaria) and euglenoids (Euglena). reinhardtii. 2005) and control translation by either 1950s (Bird and Benson. 1987).. 2001). vortexing with glass beads high throughput transcriptomics (i. (Kindle et al. Porphyra).. Dunahay. DNA introduced into the nucleus sense. which can then subsequently affect algae algal transgenics and implications for biotechnology. small sizes and used to target suppression of specific and traditional mutagenesis and breeding has been used genes by base pairing. including conjugation. Gene Expression Control Elements Porphyridium.. need for balance in cellular metabolism. brown algae Gene expression control elements (also known as (Laminaria. a fundamental can be actuated by simple manipulations are desirable. 1995). 18 2.. One solution to this problem was developed in dominance issues when more than one copy of the C. and so recessive genotypes or when dealing with redundant genes where traditional may not manifest unless that genotype is allowed to genetic manipulations are difficult. 1991) or silicon carbide whiskers (Dunahay. Control element strength can be evaluated Other successful methods include electroporation by monitoring mRNA levels by quantitative PCR or (Shimogawara et al. Even on vectors containing both the genome (Dunahay et al. because it is induced could be applied. be desirable.. 2004). Gene replacement a selectable marker and an RNAi construct. inducible and repressible promoters that 1993). then chemically based methods of transformation useful in this regard in microalgae. and repressed with for cyanobacteria. red algae (Cyanidioschyzon. 2006).. 1988. Many transformation methods also exist with nitrate in the growth medium. classical genetic approaches using from the ribosome (Valencia-Sanchez et al. Chlamydomonas reinhardtii. 1993). Sexual Crossing Breeding of desired characteristics from a number of RNA Interference (RNAi) phenotypic variants can allow for strain development RNAi can be a useful tool to down-regulate gene without creating GE algae. If methods exist to remove or perforate the expression. 1992). Frequently. In addition.. 1995). or reduced levels of expression may transformation (Boynton et al. Algal Biology  . 2005). transcriptional regulators) can modulate the levels Hallmann (2007) provides a comprehensive review of of mRNA. Identification of and biolistic approaches (Matsunaga and Takeyama. RNAi operates “breed true” though a series of sexual crosses. (Deng and Capecchi. traits. electroporation. and can be used to knockout genes. or co-introduction of recombinase genes with the transforming DNA (Reiss et al.

. have enabled in-depth analyses to be performed in a whole cell context. with a special focus on classes or individual High throughput approaches.. at least semi-quantitative evaluation of their abundance assembly of such data (especially with short read lengths) in a simple measurement. which could impact the regulation Enabling Technologies: “Omics” and processes of fuel precursor production. Sequenced genomes are an essential basis of information bioinformatics can represent the largest stumbling block for the interpretation of transcriptomic and proteomic to achieving that goal. A genomic survey of representatives from all major algal classes is Approaches and Bioinformatics desirable. Even though new sequencing technologies determining their intracellular location and can provide readily enable accumulation of data for large Department of Energy) and for Ectocarpus siliculosus useful for applications. Osteococcus tauris.html 2. 2006). 2008). Although time families). Genome size in algae can vary substantially. for which over 20 completed these methods have revolutionized the study of organisms genome sequences are available. With the development of more powerful sequencing training should be established at the onset of activities methods.).Directed Evolution of Enzymes/Proteins reduced and more coverage is obtained in a shorter period of time. and proteomics. transcriptomics. a pelagophyte (Aureococcus annophageferrens). applied research and technology development. even in closely related species (Connolly et al. Algal Biology  19 . demonstrated utility and some possible advantages It is likely that the different symbioses have affected the (Regoes and Hehl. Together. creating the new field of bioinformatics unicellular green algae (C. the macroalga Porphyra purpurea at the Joint Genome Institute characterization of these cellular processes will prove (U.Centre National de Séquençage (France). pussilla. a coccolithophore (Emiliania huxleyi). approaches to better understand the underlying Thalassiosira pseudonana. especially at the basic level of gene annotation. Bathycoccus sp. Volvox carteri). cellular processes and regulation involved in Genome sequencing projects are also underway for the defining the attributes of the strain..1 These species include computer sciences. a be strongly considered for any strains being developed substantial amount of regulation occurs at the protein for biofuels research or production. Gaietta et al. such as genomics. useful in monitoring intracellular metabolic processes associated with biofuel precursor production. all featuring a complex series of primary widely used and versatile protein tags. a where powerful new databases and search algorithms are red alga (Cyanidioschizon merolae). so it will be important to provide sufficient consuming. will be essential Sequencing and Annotating Algal Genomes to make sequence data usable. species within classes that make abundant fuel precursors. Green Eukaryotic algae constitute members from at least eight fluorescent protein and its derivatives are the most major phyla. This could 1 For a listing. 2002).. that the genomic data are only as useful as feedback. but others have and secondary endosymbioses (Falkowski et al.jgi-psf. Algal species are being analyzed using these analytical and several diatoms (Phaeodactylum tricornutum. These biological of only a handful of microalgal species have been fully advancements have been complemented by developments in or partially sequenced to date. approaches to modify proteins by genetic resources to allow for detailed analysis of the data. Fragilariopsis cylindrus). Except for cyanobacteria. 2006). Quality standards and appropriate data. It must be noted level. engineering so that they function more efficiently or have other favorable characteristics could be valuable for the development of algal biofuels technology. the nuclear genomes both in culture and in natural habitats. reinhardtii. This information could be can be more challenging in repeat-laden genomes. obtaining a genome sequence should Regarding core cellular metabolic processes. One reason for this variation is likely to be the accumulation Protein Tagging Technologies of repeated sequences in the larger genomes (Hawkins Tagging proteins with fluorescent markers is useful in et al. distribution of DNA between the plastid and nucleus (Wilhelm et al. in which costs have been substantially to ensure consistent and useful annotation. 2005.S. including allosteric activation and metabolic though. If not properly done. Bioinformatics analysis of sequenced genomes. this level of regulation integrates the annotation (the assignment of gene functions or the proteome with the metabolome. Micromonas in ways and timescales that were never before possible. forming the foundation for at Genoscope . Indeed. several picoeukaryotes helping biologists share and build upon experimental results (Osteococcus lucimarinus.. 2004). visit http://genome. Undoubtedly.

Dettmer et al. spontaneous considered in investigations of gene expression regulation. different metabolomic analysis tools are applied. Because the chemical that carry out similar functions can also help assign gene nature of metabolites varies more than for mRNA function and identify metabolic pathways of interest. identification. Most transcriptomics approaches Cyanobacteria evaluate mRNA levels. but they are prolific carbohydrate and secondary approaches allow for robust evaluation of soluble and metabolite producers. however. response of living systems to pathophysiological such as Dunaliella sp. Algal Biology  . 1999). and proteins. and function. as well as spectrometry. Scenedesmus stimuli or genetic modification (Nicholson et al. 2006). Both metabolomics and metabonomics may be important in terms of algal biofuels research. Lipid analysis is done using mass spectrometry of any algal biofuels technology development effort. These approaches not only enable protein atmospheric nitrogen and produce hydrogen. another dozen or so EST projects. was sequenced over a decade ago. notable that while these projects represent a very useful There is a distinction between metabolomics. sp. but also allow for protein quantification and many can be genetically manipulated. double-homologous recombination. are underway. 2007). at least10 other algal genomes.. Mass-spectrometry lipids. 2009). 2007) and should be applicable in cyanobacteria. The cellular complement of proteins reflects its metabolic potential. in response to particular environmental conditions. 2005. Chlorococcum sp... Cerutti more advanced than that of eukaryotic algae because and Casas-Mollano. 2008). 2007. Tanner et al... For example. Transcriptomics Lipids are a subset of the molecular repertoire of the While genome sequencing will be an important component algae cell. Comparative genomics weight compounds in a cell that are involved in growth. Applications of Biotechnology New. 2004. transformation. small RNAs also play Genetic manipulation of cyanobacteria is generally major regulatory roles in algae (Bartel. and ultimately determines how a cell functions Cyanobacteria generally do not accumulate storage in response to the environment.. and metabonomics. high-throughput sequencing technologies enable to Algal Bioenergy comprehensive coverage of transcripts and quantification of their relative abundance.. gas chromatography mass spectrometry. Castellana et al. or can determine fuels and intermediates. It is and nuclear magnetic resonance (Dunn et al.. 2005). Monitoring lipid characteristics under a coding regions of DNA).. Small RNAs have been many of the tools developed for bacterial genetics are identified in microalgae (Zhao et al. the rationale for sequencing these organisms was involves the identification and analysis of metabolites. identifying final fuel produced. and at least half a dozen selectable markers Proteomics are available for Synechocystis (Vermaas. but it is also emerging as a variety of different cultivation regimes can also help inform robust approach for genome-wide expression analysis process engineering and improve yields (Yu et al. making them detection of post-translational modifications (Domon and attractive organisms for biofuels production.. proteomic Synechocystis has been used extensively for the study of approaches can also help assess the metabolic potential of carbon metabolism toward production of hydrocarbon organisms that may be difficult to isolate. In the absence of genomic information. 2006. which survey. which is the quantitative Therefore. Moreover.include the standardization of using a particular sequencing Metabolomics and Lipidomics approach that provides sufficient coverage of ESTs to The metabolome is the collection of small molecular ensure accurate gene modeling.. quantitative transcriptome profiling using new. Many 20 2. Some strains can double membrane-associated proteins in the form of protein quickly (less than 10 hours). approaches between related organisms and organisms maintenance. including liquid chromatography mass Currently. Aebersold. Nannochloropsis sp. approaches (Han and Gross. The genome of this strain functional diversity of a community of organisms. not related to lipid production or other biofuels efforts.. the first among photosynthetic organisms (Kaneko et al. and protein tagging are routine in some cyanobacterial systems. 1996). high Quantitative comparison of lipid type and abundance are throughput sequencing technologies will also become critical components of lipid-based biofuels approaches as increasingly important because it will not only help lipid characteristics can determine the suitability of the with genome annotation (for example. 1998). especially with regard to translational regulation. and Pseudochlorococcum sp. and some strains can fix peptides. there are still a large number of useful algal measurement of the dynamic multi-parametric metabolic species to sequence that are related to biofuels production.

reinhardtii has been recently been (sinks) in cyanobacteria. 2006). 2008). and isobutanol (Atsumi lipid producer. In order to redirect carbon to a fuel al. These studies can not only serve to advance examples of Chlorella transformation have been reported. C. the understanding of how the production of different although the stability of the expression of the foreign carbon storage molecules are controlled in response to genes is still questionable (Leon and Fernandez. It has not been shown how these dry weight lipid (Xu et al. It produces abundant Microalgae lipids (Weldy and Huesemann. 2009.. Chlamydomonas reinhardtii is the most studied eukaryotic algae. and events likely had significant effects on the metabolic transgenic strains have been reported (Li et al. physiological conditions. and because it has outstanding salt tolerance (from 0... anaerobically (important for the production of hydrogen). A possible drawback of C. and polyhydroxybutyrate Chloroplast transformants are also stable. 2007).html 2. which occurs in the Diatoms were a major focus in the Aquatic Species Program chloroplast. Dunaliella salina has several useful characteristics for large-scale biofuels production. 2004).1 M to near Unicellular eukaryotic microalgae are the product of saturation). 2006. et al. it will be necessary to further being developed. 2006). it can be grown under extreme conditions over 3 billion years of evolution. protothecoides. 2007 ). 2003). In can accumulate under salt stress (Yoo et al. For example. sucrose (Nobles and Brown. like the production potential (Tamagnini et al.. Ferrante et al. and metabolome (Singh et al. and positional effects (Ferrante et in all cyanobacteria. 2009) in Synechococcus.. 2008).. 2000). capabilities. In addition to having a sequenced nuclear genome Transgenic approaches have enabled the production and (Merchant et al. which are cells is also often dependent on environmental conditions. accumulation. including glycogen. Koksharova et al..jgi-psf. C. and chloroplast (PHB). Relatedly. but nevertheless. 2007. resulting in greater than 50% (Miyake et al. However. 1999).. Continued exploration of the evolutionary A distinguishing feature of diatoms is their silica cell walls... and high-throughput analytical techniques are adept at making fuel precursors and with high have been applied to the study of its transcriptome.home. strategies to address these issues are production pathway. 1998). algae with different extents of endosymbiosis (Wilhelm suggesting favorable growth rates for biomass production. filamentous strain that can form heterocysts. 2007). proteome.. is at least partly regulated by nuclear-encoded given their ability to accumulate high amounts of lipids gene products. fatty acid synthesis. a comprehensive understanding due to codon bias (Heitzer et al. For example. Fulda et al. Algal Biology  21 . it can be sexually crossed. Multiple endosymbiotic events organisms. It has also been shown that induces heterotrophic growth.. 2007). productivity under various environmental conditions. The latter is a production of other carbohydrate-based storage compounds. Eisenhut et al. Lipid production. reinhardtii can serve et al.2 Several production. the addition of an external carbon source Xiaoling Miao et al. 2007) and well developed transgenic secretion of cellulose.. 2002). reported (Neupert et al. 2006. pathways and regulation of fuel precursor synthesis. It is not an abundant ethanol (Deng and Coleman.. 2008).. salina is occurred during the evolution of microalgae.. with specialized structure and metabolism that function some of which await elucidation and development. but may also serve to guide the development of other types of algae for biofuels production. 2004).photosynthesis and carbon metabolism mutants have been diversity of algae is important to identify species that generated. glucosylglycerol. 2008. it is known that glycogen accumulates under normal growth conditions in Chlorella is another well-studied genus of green algae. These currently being determined (estimated size 130 Mb). The genome sequence of D. Diatoms are responsible for 20% the interaction between the nucleus and chloroplast in of the total global carbon fixation (Armbrust et al. reinhardtii is the fact that nuclear expression of foreign genes is still problematic Despite all of the progress. Synechococcus as a model system for understanding the fundamentals of and Anabaena strains have been studied for their hydrogen lipid synthesis and regulation. and the conditions that trigger carbohydrate protein expression systems are well developed. RNA silencing of carbon metabolism and regulation is not yet available (Cerutti et al. and there are fundamental differences in (Sheehan et al. 1997).org/ChlNC64A_1/ChlNC64A_1. whereas glucosylglycerol and sucrose and some species are abundant lipid producers. and stable and successful expression characterize the dominant carbon storage compounds of foreign genes in C. sucrose.. which increases both growth PHB accumulates under nitrogen depleting conditions rate and lipid production. The genome sequence pathways can be manipulated for the benefit of biofuels of Chlorella NC64A was recently completed... Synechocystis. 2 http://genome. 2008).. and are highly diverse that could reduce the growth of possible contaminating (Falkowski et al..

. there have been several unintended releases (red algae) on the basis of their predominant pigments. Niwa et al. and regulatory gene expression control 2005).gov 22 2. Recombination of and possibly with homologous recombination. 1999. 1997. They have low lipid content as a engineering of seaweed genes to create valuable products. and four more 1999. being applied to understand cell function in representatives With the development of robust gene silencing approaches of red. Green macroalga such as Ulva (also known as sea lettuce) are classified together with common green microalgae such as Chlamydomonas and Chlorella as Chlorophyta.. which includes diatoms. For the species displaying variations of annual and perennial purpose of this report. with different the appropriateness of deploying these strains. Niwa et al. 2003).. GE algae are defined as strains life histories. For example... are very important. potential for large-scale blooms and subsequent is in a state of flux (Ali et al. Bouza et al. or seaweeds. Zaslavskaia (Jacobsen et al. and strain selection and been focused explicitly on biofuels.. Bowler et al.3 None of the sequencing projects has thus far et al.and their requirement for silicon as a nutrient for growth. et al.. and alternation of generations. The life cycles a great deal of uncertainty regarding the need for or of macroalgae are complex and diverse. taxonomic affinities are under re-examination 2008).. 2008). and green seaweeds. the brown macroalgae such as the kelps are GE algae as production strains. They are and proteomic profiling. 2005). 2005). 2006) are elements have been identified (Poulsen and Kroger. induction response.doe. and holdfast for anchoring the entire structure to Despite the great promise of GE algae. the gene existing genes through selection and procedures such manipulation toolkit for diatoms will be fairly complete. represent a broad group of projects will facilitate efforts such as global genomic eukaryotic photosynthetic marine organisms. each of the major macroalgal groups has in terms of organism containment. of GE organisms over the past 15+ years (GAO report. 2005). Modifications They are typically comprised of a blade or lamina. Currently. Apt et al. 3 http://www. also have microalgal counterparts. are well established for several diatom species (Dunahay et 1999. 2000). Moulin et al. Two diatom genome sequences are complete are used in seaweed population genetics (Alberto et al. 2003. development of genetic classified as Heterokonta within the Chromalveolata. 2006. (Armbrust et al.. constructing detailed pathways evolutionarily diverse and abundant in the world’s oceans for secondary metabolite production. 1995. Algal Biology  . 1994. 1996. anoxic zone formation. affinity with corresponding microalgal forms. Meneses and Santelices. 1995. Baldauf. Use of gene-specific probes al. there have usually been built in safeguards to prevent the release of GE organisms to avoid potential Macroalgae are historically classified as Phaeophyta disruption of ecosystems. Understanding the basic biology that will inform with the use of molecular tools and phylogenetic such aspects as lateral gene transfer... 2001. Red macroalgae such as Porphyra spp.. general rule but are high in carbohydrates that can be converted to various fuels. potential for toxin markers. as protoplast fusion will be the basis for new strain creation where outplanting of individuals for growth Macroalgae in natural environments is a goal. Transgenic techniques characterization (Jin et al. species. tools is still imperative from a research standpoint. Unlike microalgae. As such. For Despite the uncertainty regarding the development of example. Ho are underway. such as the unicellular alga Porphyridium cruentum.. and expression profiling (Collen et al. the stipe. 1999.. 2004. Chlorophyta (green algae). there is nevertheless hard substrates in marine environments. because silicon metabolism is not restriction fragment length polymorphisms (RFLPs) and believed to be tightly coupled with the metabolism of other random amplified polymorphic DNA (RAPD) analysis nutrients. brown. and choice of cultivation methods In general. Fischer et al. Dutcher and Kapraun. Roeder et al. and metabolic and coastal waters.jgi. This is advantageous for studying the lipid of seaweed genetics and cell function. even with these (brown algae).. Since the beginning of the deployment of GE organisms.. Advances in seaweed cell and molecular biology are Silicon limitation is one trigger for lipid accumulation currently being applied toward a better understanding in diatoms. However. Genome sequencing Macroalgae. the status of macroalgal systematics production. they are Considerations of Genetic multicellular and possess plant-like structural features. and Rhodophyta safeguards. combinations of sexual and asexual carrying coding sequences obtained from a foreign reproductive strategies..

304-311. 306(5693). Boynton. Libourel. R.E. E. H.A. On the origin and functions of RNA-mediated silencing: from protists to man. S. Arabolaza. CDlNp&page=1&doc=3. BMC Systems Biology. Boyle. Y. Metabolic flux analysis in plants: coping with complexity. Cell & Environment. (2005).C. 63(12). Klein.Gramajo. C. mechanism. E. et al. Moroney. et al.. Kawachi. (1988). The life cycle of neutral lipids: Synthesis. Elsevier. 16(5). Bird. (2004). Johnston.. 51(3).P. (2008). (2006).. evolution. (2009). S. (2005). Armbrust. Proceedings of the National Academy of Sciences. 60(4). D.G. Elsevier Science Ltd. Nature Biotechnology. 21034-21038. 83–102. Phylogenetic relationships among algae based on complete large-subunit rRNA sequences. biogenesis. et al. Science. J. Dyer. Chloroplast transformation in chlamydomonas with high velocity microprojectiles.C. Andersen.A. Amsterdam. H. K. 2010. Assessing patterns of geographic dispersal of gelidium sesquipedale (rhodophyta) through RAPD differentiation of populations. Progress in Lipid Research. P. (1987). W. S. et al. 50(2). Ceron Garcia.. rhodophyta) investigated by random amplified polymorphic dna (rapd) markers. Benning. Ynalvez.. Cell. et al. G. Molecular and General Genetics MGG. Algal Biology  23 . storage and degradation. J. 737-749. Daum. 456(7219).G.V. E. (32).. Current genetics. Cerutti.E. 1241-1257. Bowler. Ye. Traditional microalgae isolation techniques. Archiv für Hydrobiologie. (2009). Botryococcus braunii: A renewable source of hydrocarbons and other chemicals. Progress series(Halstenbek) (191). (2009). Multiple pathways for triacylglycerol biosynthesis in Streptomyces coelicolor. 367-376. S. Triacylglycerols in prokaryotic microorganisms. Steinbuchel. & Merino. (2008). Alberto. 3. Epigenetic silencing of a foreign gene in nuclear transformants of chlamydomonas.L. Biolistic nuclear transformation of Saccharomyces cerevisiae and other fungi. Mason.M.References Adams. K. Apt. I. Applied Microbiology and Biotechnology. 9(6). (2006). J. J Morgan.T. 22(3). 245–279. Bouza. 81–99. Ahlgren. and metabolism. (2002). The Genome of the diatom thalassiosira pseudonana: Ecology. et al. Allen.C. Mixotrophic production of marine microalga phaeodactylum tricornutum on various carbon sources.. Leitao. Baldauf. Lipid analysis of freshwater microalgae: a method study. 252(5).. Santos. and function. (1999). 1991. Science. F. 105(52). Tural. et al. Retrived January 5.. et al. Sanford.. J.A.J. Ben Ali. (1990).V. (2008). Bartel. 47(5). Applied and Environmental Microbiology. Canadian Journal of Physiology and Pharmacology. Plant. K. (2001).. Journal of Phycology. R. A rapid method of total lipid extraction and purification. Nature. Algal Culturing Techniques. W. Functional Plant Biology. Altabe.E. Cerutti. A. Seaweed Cultivation for Renewable Resources. K. Genetic structure of natural populations in the red algae gelidium canaariense (gelidaiales. 101–108. 121(3). D. R. The Phaeodactylum genome reveals the evolutionary history of diatom genomes. Stable nuclear transformation of the diatomphaeodactylum tricornutum. 689–694.isiknowledge. Casas-Mollano. Discovery and revision of arabidopsis genes by proteogenomics. N. Critical reviews in biotechnology. (1996). M. Higashide.. 1177-1180. A role for lipid trafficking in chloroplast biogenesis. 42(2).M. Current genetics. The Plant Cell Online. Rodriguez. H. A. D. J.K. B. PH. 911–917. (1959).L. International Journal of Systematic and Evolutionary Microbiology. A. (2003). (2006).B. H... Alvarez.74(9). 300(5626). Kroth-Pancic.. (2008). Cellular and Molecular Life Sciences: CMLS. A. 32(9). Castellana. (1997).E. C. Direct photosynthetic recycling of carbon dioxide to isobutyraldehyde. Armaleo. A mutant of chlamydomonas reinhardtii that cannot acclimate to low CO2 conditions has an insertion in the Hdh1 gene. Journal of microbiology and biotechnology. Bligh. Shachar-Hill. Athenstaedt.. 239-244. (2006). 55–66.B. J. Flux balance analysis of primary metabolism in chlamydomonas reinhardtii. L. 1703-1706.L. 17(2). S. M.. 240(4858). 572–579. 116(2). T. 97–103. Grossman. http://apps. MicroRNAs genomics. N. H.. Liao. Science. Colombo. 381–389. The deep roots of eukaryotes. Banerjee. C. N. G. 2573-2582. (2002).R. 295–306. 2. 27(12). 281–297. Shark. G. Benson. J. et al. (2004). 925-945. A. 79-86.G. 37(8). 1534-1538. Marine ecology. Atsumi.

K. A. The EMBO Journal. et al. 849–857. 65(2). F. (1992). Norazmi Mohamed. 463–515.R. Phospholipid: Diacylglycerol acyltransferase: an enzyme that catalyzes the acyl-CoA-independent formation of triacylglycerol in yeast and plants. Journal of Cell Science. Roessler. Fuhrmann. 3(9). Biodiesel from microalgae. (2007). 15(5). P. Measuring the metabolome: Current analytical technologies. et al. (2008). Finazzi. Ferrante. (2007). 305(5682). et al. Purton. Genetic transformation of the diatoms cyclotella cryptica and Navicula saprophila. 2803-2809. 21-27. 354-360. Ethanol synthesis by genetic engineering in cyanobacteria. Journal of Phycology. Malaysian Journal of Science. (1989). Eberhard. (1971). (2003). M. (1999).D. (2005). (1995). 22(2). A. 212-217.A. T. Biotechnology Advances. Random amplified polymorphic DNA (RAPD) identification of genetic variation in three species of porphyra (bangiales. S. 312(5771). (2006). Connolly. Photooxidative stress in plants. Rochaix. S. N. Journal of Microbiological Methods. E. (2000). M. (2007). Dunn. J. Proteome analysis of salt stress response in the cyanobacteriumsynechocystis sp. 606–625. Finding gas diffusion pathways in proteins: Application to O2 and H2 transport in CpI [FeFe]-hydrogenase and the role of packing defects. 639-642.. Kapraun. Debuchy. 6487-6492.G. 3365-3371. strain PCC 6803.. Cohen. Del Pozo. et al. (2008). Metabolome phenotyping of inorganic carbon limitation in cells of the wild type and photorespiratory mutants of the cyanobacterium synechocystis sp strain PCC 6803.G. Molecular and Cellular Biology.. Expression profiling of chondrus crispus (rhodophyta) after exposure to methyl jasmonate.J. 523528. J. M. 25(3). B. European Journal of Biochemistry. D.105(1). 452. Wollman. Journal of Phycology. 267-273. 124–131. Plant Physiology. et al. Collen. J. 97(12). R.. Fluorescent measurement of microalgal neutral lipids.. Applied and Environmental Microbiology. 6(6). rhodophyta). Structure. The Analyst. 113–120.. (1994). S. 12(8). Eisenhut. e3200. Chisti. De la Jara. An optimized. J. 6(9). R. Hammock. Fulda. The evolution of modern eukaryotic phytoplankton.E. Prometrics. 2733-2745. 68(3). (1999). Dunahay. (1994). 31(6). nostoc muscorum. Two different genes from schwanniomyces occidentalis determine ribosomal resistance to cycloheximide. Targeting and covalent modification of cell wall and membrane proteins heterologously expressed in the diatom cylindrotheca fusiformis (Bacillariophyceae). J. 42. 92(4). (1987). et al. L. (2008). G. Fatty Acid Composition of Some Malaysian Seaweeds. C. 26(1). L. Dahlqvist.R. Annual Review of Genetics. Mass spectrometry and protein analysis. Flow cytometric determination of lipid content in a marine dinoflagellate. 3857-3863. 15(3). B. 433–438. Bowen.D. C. (2005). & Phang.G.D. Biotechniques. et al. Coleman. chemically regulated gene expression system for Chlamydomonas. Correlated evolution of genome size and cell volume in diatoms (bacillariophyceae). W. The argininosuccinate lyase gene of chlamydomonas reinhardtii: An important tool for nuclear transformation and for correlating the genetic and molecular maps of the ARG7 locus. H. Mass spectrometry reviews. (2003). 171. 696-717. Algal Biology  . D. The abundant retinal protein of the chlamydomonas eye is not the photoreceptor for phototaxis and photophobic responses. W. Journal of Phycology. (2006). crypthecodinium cohnii. (2004). Purification and properties of glycogen isolated from a blue-Green Alga. Domon. K. J. Capecchi. Fischer. M. 35(1). Falkowski.. Chu. et al. 13(9). P. Reexamination of gene targeting frequency as a function of the extent of homology between the targeting vector and the target locus. Foyer. Science.H. et al. Journal of Applied Phycology. et al.E. 51-78. 130(5). 1321–1329. Kunert. 148(4). Deng. Jarvis. Physiologia Plantarum. Bailey. 6(3). et al. 44(1).Chao. 2109-2120. 213(2). et al. C. (2001). Elsey. et al. 8(10). Dutcher. 294–306. (1993). Proceedings of the National Academy of Sciences. (2006).C.. PloS One.. Y. Cooksey. Mass spectrometry-based metabolomics. P. 114(Pt 21). 333–345..E. 1004–1011. Journal of Bacteriology. Deng. Johnson. Aebersold.G. Journal of Experimental Botany. 24 2. Lelandais. Aronov.A. Fluorometric determination of the neutral lipid content of microalgal cells using nile red. Transformation of chlamydomonas reinhardtii with silicon carbide whiskers. M. K. The dynamics of photosynthesis. T. 331-338. (1993).A. Dettmer. Science Signaling.B. P. Journal of Applied Phycology.J. H. (2008). et al. S. Dunahay. Journal of Microbiological Methods.

.. E. 616. Annual Review of Plant Physiology and Plant Molecular Biology.S. Lüning. (2003). Rasmussen. F. Hu. Horn. J. 6-53. PA and Murata. Influence of codon bias on the expression of foreign genes in microalgae. I. 296(5567). 621–639. II. M. et al. 1-11. (2000). E.). 99–110. Advances in Experimental Medicine and Biology.doi. Krause. 249-254. Retrieved December 30. Journal of Cell Biology. 109-136. Nuclear transformation of eukaryotic microalgae: historical overview. Weis. N. et al. K. (2007). 75(1). Hawkins. Plant Physiol. 81-98. 9(6). Østgaard. 71-91. 253–264. Leung. DNA-mediated transformation of chlamydomonas reinhardi cells: Use of aminoglycoside 3’-phosphotransferase as a selectable marker. 182(5). W. (2007). 58. The Netherlands: Kluwer Academic Publishers. 39(5).C. Koksharova. et al. Expert Review of Proteomics. Hoiczyk. K. analysis of fermentative products from starch in dark and light. O. León. 25(5).E. Shotgun lipidomics: Multidimensional MS analysis of cellular lipidomes. Chlorophyll fluorescence and photosynthesis: The basics. Genome Research. 1(1). S. Stable nuclear transformation of chlamydomonas using the chlamydomonas gene for nitrate reductase. Differential lineage-specific amplification of transposable elements is responsible for genome size variation in gossypium. 795–805. et al. (2000). (2007). T.L. Journal of Phycology. Huang. (1997). C. A. Multicolor and electron microscopic imaging of connexin trafficking.M. (2007). A. 313-349. Algal Biology  25 . Hallmann. (1995). G. Dordrecht. Journal of Applied Phycology.1007/978-0-387-75532-8_5. 212-218. 1721-1725.K. Rappel.. Goulard. Manavathu. Circadian changes in relative abundance of two photosynthetic transcripts in the maring macroalga kappaphycus alvarezii (rhodophyta). et al. Proceedings of the National Academy of Sciences.L. J. et al.J. The Plant Journal. 7(1). 3(3). G. Gibbs. Annual Review Plant Biology. 54(4). strain PCC 7942. Jacobsen. 5(12). 2009. S. Ethanol production from seaweed extract. Aasen. X. (1984). 16(10). eds).. & Fernández. 403(1- 2). 42(1). In transgenic microalgae as green cell factories. The Journal of Bacteriology. 88(5). Kindle. Hallmann. (1991). function and genetics (Siegenthaler. Production of ethanol from mannitol by zymobacter palmae. The nitrate reductase gene-switch: A system for regulated expression in transformed cells of dunaliella salina. et al. Ho. D. Pang. (1998). Kindle. strain PCC6803. Molecular and Cellular Biology. F. 489-493. (1992). M. U. K.L. S. 1252-1261.W. Fermentative metabolism of chlamydomonas reinhardtii: I.. Plant journal. The first protein map of synechococcus sp. (1989). S. (2002). Heitzer. Richards. J. 132-142. Molecular characterisation of sargassum polycystum and S. Hydrogenases and hydrogen photoproduction in oxygenic photosynthetic organisms*. R.B. J. Hansel.. Ghirardi. (1991).Gaietta. Sequence determination of the entire genome and assignment of potential protein-coding regions. 503-507. et al. DNA Research. 177-200. 888-896. Han. Gfeller. A. Gene. (2007). A. Rawsthorne. E.A. Horn. Østgaard. Kang. et al. 43(1). 17. K. Hasnain. 53–64... R. Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. http://dx.. I.L. Stern. 664-672. Jin. 75(6). R. 3647-3650. Li. (1994). S. Cyanobacterial cell walls: News from an unusual prokaryotic envelope. Kaneko. Membrane lipids in algae.. Starch and fatty acid synthesis in plastids from developing embryos of oilseed rape (Brassica napus L. H. 33-41.. Journal of Industrial Microbiology and Biotechnology. 2.M. Harwood. A. (2006). S. Journal of Industrial Microbiology and Biotechnology. siliquosum (Phaeophyta) by polymerase chain reaction (PCR) using random amplified polymorphic DNA (RAPD) primers. 24(1). Engineering the chloroplast genome: Techniques and capabilities for chloroplast transformation in chlamydomonas reinhardtii. 51-57.H. (2000).L. (2005).P. Annual Review of Plant Physiology and Plant Molecular Biology. Science. 2(2).C.H. 109(6). (1985). Gross. Algal transgenics and biotechnology. K. Oil bodies and oleosins in seeds. Q. (2008). Aasen. Plant Journal. achievements and problems. Phang. K. E. Journal of Applied Phycology. Klint. 2589-2601. (1996).J.. M. Genetic engineering of the multicellular green alga volvox: A modified and multiplied bacterial antibiotic resistance gene as a dominant selectable marker. T. Transgenic Plant Journal. (2006). 1191- 1199. (1999). 6(6).. Lipids in photosynthesis: Structure. Gelling agents for tissue culture of the seaweed hizikia fusiformis. Microalgal triacylglycerols as feedstocks for biofuel production: Perspectives and advances.

B. Kikuchi.K. K. et al. P. 15(6).11(3). International Journal of Hydrogen Energy. Meneses. 77-84.M.. Progress in lipid research. 50(1). (2005). C. Morphological and molecular analysis of the endangered species porphyra tenera (bangiales. Generation of chlamydomonas strains that efficiently express nuclear transgenes. G. Xenobiotica. K. Molecular and Cellular Biology. Ohlrogge. Algal Biology  . D. Annual Review of Plant Biology. (1994). Polyhydroxybutyrate production from carbon dioxide by cyanobacteria. Kobiyama. Metabolic flux analysis in plants: From intelligent design to rational engineering. 34(17). Strain selection and genetic variation in gracilaria chilensis (gracilariales. 245- 250.S. 333-359. Niwa. Journal of Phycology. (2000). J.. P.K. S. The biogenesis and functions of lipid bodies in animals. (1999). J.. rhodophyta). M. Journal of Applied Phycology. FEMS Microbiology Letters. The complex network of non-cellulosic carbohydrate metabolism. N. ‘Metabonomics’: Understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. Sucrose accumulation in salt-stressed cells of agp gene deletion-mutant in cyanobacterium synechocystis sp PCC 6803. Wang. et al. Nutritional composition of edible Hawaiian seaweeds. Fernie.J. 53(4). Current opinion in plant biology. Cellulose. I. J. Merchant.. 957-970. 45(1). (2006). Progress in Lipid Research. Annual Review of Plant Physiology and Plant Molecular Biology. Lipid biosynthesis.. Miyake. Shachar-Hill. (2007). Diacylglycerol acyltransferase: A key mediator of plant triacylglycerol synthesis. 325–438. R. B. N. Savereide. (1999). (2007). Santelices. (2003). Holmes. Miao... A. a volvox gene required for asymmetric division and germ cell specification. rhodophyta) by polymerase chain reaction restriction fragment length polymorphism analyses of the plastid and nuclear DNA. K. Journal of Applied Phycology.P. P. Takeyama. J. Y.L. 59. 71-77. Y. 40(5). Lipids. 991-1002. (2000). T. Nicholson.A. Lytovchenko. A. Aruga.G. 1073-1088. & Zhao. function and biogenesis of storage lipid bodies and oleosins in plants. (1999). The Plant Journal: For Cell and Molecular Biology.G. E.. Metabolic pathway engineering for enhanced biohydrogen production. (2005). Aruga. 691–701. 122(1).J. Yocum. Murphy. Brown. McDermid. X. J. 1237-1245. Miao. Nelson. D. Wu. Weselake. (2001). The Chlamydomonas genome reveals the evolution of key animal and plant functions. 57(6). et al. (2003). 218(1). Miller. B. 521-565.. Lung. Nelson. (1995). Y. Photoinhibition of photosynthesis in nature. 241-246. Annual Review of Plant Biology. J... Confirmation of cultivated porphyra tenera (bangiales. Plant Physiology. (1994).C.. Isolation and characterization of six cDNAs involved in carbon matabloism in laminaria digitata (phaeophyceae).R. glsA. Lefebvre. Niyogi. D. 1181–1189. I. Wu. 127-135. (2009). Q. D. 633-662. S. 296-302. 41(12). Falkowski. encodes a chaperone-like protein. Niwa. The CRY1 gene in chlamydomonas reinhardtii: Structure and use as a dominant selectable marker for nuclear transformation. 318(5848). 218(1). Mathews. 15(5). 4011-4019. Sustained photobiological hydrogen gas production upon reversible inactivation of oxygen evolution in the green alga chlamydomonas reinhardtii. J. U. plants and microorganisms. 625-650. Melis. A. 247- 280. S. 126(4). et al. (2008). Science..A. S. The Plant Cell. (2008). 41(2). K.J. Browse. (1999). Genetic engineering in marine cyanobacteria. Bock. 84- 86(1).. (2006). Annual Review of Plant Physiology and Plant Molecular Biology. Neupert. P. D. Karcher. 10(3). 7404-7416. Nobles. 7(1). G. 513-524. Journal of Phycology. & Stuercke. S. 35.. 294-304. 14(6). Transgenic expression of gluconacetobacter xylinus strain ATCC 53582 cellulose synthase genes in the cyanobacterium synechococcus leopoliensis strain UTCC 100. Applied Biochemistry and Biotechnology. Structure and function of photosystems I and II. Photoprotection revisited: Genetic and molecular approaches. (2003). et al. (2009). Structure. N. 29(11). 1140-1150. R. (1993). Murphy. FEMS Microbiology Letters.. (1995).L Kirk. Humphries. Sonnewald.. 7(7).. 71-77. A.. Lindon. Development. J. Journal of Applied Phycology.R. Sucrose accumulation in salt-stressed cells of agp gene deletion-mutant in cyanobacterium Synechocystis sp PCC 6803. 32(3).M. (1999). X. 227–235. Moulin. H. Long. 57. Phycological Research. 649-658. R. 26 2. Matsunaga. rhodophyta).Libourel.

273-291. 40(4). et al.J. 236(1). (2005).G. et al. Y. S. Automated isolation techniques for microalgae. et al. Induced beta-carotene synthesis driven by triacylglycerol deposition in the unicellular alga dunaliella bardawil. RecA protein stimulates homologous recombination in plants. Wassarman.. Shachar-Hill. M. et al.. Worth Publishers. K. et al.A. J. Kröger.. Journal of Phycology. (1988).E. T. Plant Physiology. 1191-1206. 49–59. Shimogawara.. The photobiological production of hydrogen: Potential efficiency and effectiveness as a renewable fuel. Department of Energy’s Aquatic Species Program: Biodiesel from Algae. Transient gene suppression in a red alga. Planta.. (2004). Tanaka. (2003).E. A. (2005). Academic Press.. K.. The plastidic phosphoglucomutase from Arabidopsis.M. (2008).. Kroger.B. P. Reversible hydrogenase activity of gloeocapsa alpicola in continuous culture. G. 1324-1335. Truncated chlorophyll antenna size of the photosystems—a practical method to improve microalgal productivity and hydrogen production in mass culture. An abundance of RNA regulators*. S. The Ecology of Algae. N.. 21(2).B. 179–211. Poulsen. 41(6).J. Krumbein. Stal. New York.. Identification of stress gene transcripts in laminaria digitata (phaeophyceae) protoplast cultures by expressed sequence tag analysis.R. 93(7). non-heterocystous cyanobacterium Oscillatoria sp. J.. R. et al. C. Effects of silicon deficiency on lipid composition and metabolism in the diatom cyclotella cryptica.J. Photosynthetic water-splitting for hydrogen production.. Sheehan. (1998).S. Protoplasma. Chesley. A. ASM Press.. Contrapuntal networks of gene expression during arabidopsis seed filling. 116(4).D. The Plant Cell Online. Sieracki. (2005).. (1987).. F. Reiss. K.S. Cloning and functional characterization of a phospholipid: Diacylglycerol acyltransferase from arabidopsis. Rohr. Biotechniques. 1193-1199. Melis. et al. Proceedings of the National Academy of Sciences.Ohnuma. 199-217. 217(1). H. J. Sheremetieva. Annual Review of Biochemistry. Hehl. Evert.E. P. Critical Reviews in Microbiology. C. et al. Sears. Polle. Fujiwara. 27(11-12). Singh. Plant Physiology.F. Misumi. A. Fermentation in cyanobacteria.R. 7(3).T.. 19-31. A. N. (74). 809. Crosbie. J. Tsygankov.E. International Journal of Hydrogen Energy. (2009). Kanakagiri. (1992). B. J..H. Poulton. (7). R. B. (2004). FEMS Microbiology Reviews. Tandem inverted repeat system for selection of effective transgenic RNAi strains in Chlamydomonas. Kheshgi. Riekhof. (1984). Benning. Eukaryotic Cell. S. Algal culture techniques. (2008). Storz. SNAP-tag™ mediated live cell labeling as an alternative to GFP in anaerobic organisms. 1227-1235. Stal. et al. 24. 149(1). O. 2. Regoes. 76–80.E.. (1996). 3094-3098. 467-478. Genetics. 272. N. 14(6). tla1. a DNA insertional transformant of the green alga Chlamydomonas reinhardtii with a truncated light-harvesting chlorophyll antenna size. T. Moezelaar. 148(1).M. Algal Biology  27 . 31(1). 242-52. M. Stahl. 611–621. Y. W. (2005). A reversible enzyme reaction with an important role in metabolic control. 309–317. Understanding flux in plant metabolic networks. (2006). Roessler. (2004). U. Plant Physiology. & Kuroiwa. A new molecular tool for transgenic diatoms. Altuvia. J. 107-112. 89–94. et al. High-efficiency transformation of chlamydomonas reinhardtii by electroporation. Current opinion in plant biology. Temporal separation of nitrogen fixation and photosynthesis in the filamentous. H. V. M. DC. 166(1). Ohlrogge. et al. M. Plant Physiology.. (2005). A. Journal of Phycology. Biology of Plants 5th ed. FEMS Microbiology Letters. (1998). Washington. Round.C. A Look Back at the U. 122(4). Seibert. S. (2002). Watanabe. Annotation of genes involved in glycerolipid biosynthesis in chlamydomonas reinhardtii: Discovery of the betaine lipid synthase BTA1Cr. P. N. 4(2). N. 42(5). Archives of Microbiology. Raven. (1998). Bioenergy. Eichhorn. Polle. et al. Poulsen. (1998). 148(4). 3413–3423. (1997). Ruuska. W. 1059–1065. Periappuram. The Plant Journal. L. Schwender. Serebryakova. 39(6). Integration of carbon and nitrogen metabolism with energy production is crucial to light acclimation in the cyanobacterium synechocystis. 103-116. 1239-1248. 1257–1264. The FEBS Journal. N. Molecular genetic manipulation of the diatom thalassiosira pseudonana (bacillariophyceae). S. Rabbani. (2004). L. Prince. Cyanidioschyzon merolae 10D. 135(3). (2002). (2005). (2000).E. S. 394–400. Journal of Phycology. 1821-1828. Roeder. Cambridge University Press.A. L.

Spalding. 9-24. D. deSturler. Control of translation and mRNA degradation by miRNAs and siRNAs.. High quality biodiesel production from a microalga chlorella protothecoides by heterotrophic growth in fermenters. The regulation of carbon and nutrient assimilation in diatoms is significantly different from green algae. S. Department of Energy Journal of Undergraduate Research. Yang.N..J. Metabolic Engineering. et al. Journal of Phycology. 513-526. 38(2). 4(3). Hua. J. 415-419. (126). et al. B. Huesemann. Valencia-Sanchez. 297.H. Van der Oost. 337(21-23). A. (2002). Embryo-specific reduction of ADP-Glc pyrophosphorylase leads to an inhibition of starch synthesis and a delay in oil accumulation in developing seeds of oilseed rape. Microbiology and Molecular Biology Reviews. J. S. S. Algal Biology  . 1190-1203.. Transformation of the diatom phaeodactylum tricornutum (bacillariophyceae) with a variety of selectable marker and reporter genes. et al. Lane. Journal of Biotechnology.. Journal of Phycology. International Journal of Energy Research. Wilhelm. U. Q. et al. Q. 379–386. 21(6). 17(2). (2008). A role for lipid droplets in inter-membrane lipid traffic. Nuclear-gene targeting by using single-stranded DNA avoids illegitimate DNA integration in chlamydomonas reinhardtii. Carbohydrate Research. (2005). (2009). (2000). 914-921.. 145. (2006). S. 32(5). PCC6803. Hydrogenases and hydrogen metabolism of cyanobacteria. (2008).. (1998). Microbiology and Molecular Biology Reviews. Miao. 168(1). (2007). W. (2002). 197-216. Sizova. Aristidou. P. 3904-3913. Yu. Yoo.. B. 2195-2203. Vemuri. Archives of Microbiology. Metabolic engineering in the -omics era: Elucidating and modulating regulatory networks. A complex system of small RNAs in the unicellular green alga chlamydomonas reinhardtii. Lipid production by dunaliella salina in batch culture effects of nitrogen limitation and light intensity. T. J. 293-310. Weldy. Journal of Bacteriology. 231-239. Characterization of cyanobacterial glycogen isolated from the wild type and from a mutant lacking of branching enzyme. 153–159. Zendejas. 152(5). X. 190(11). 115-122. A. Yoo. (1989).S. Renewable hydrogen production. Tanner.. 2676-2686. Metabolic flux analysis in synechocystis using isotope distribution from C-13-labeled glucose. J. 202-216. Zhu. Journal of Applied Phycology.P. (2007). strain ATCC 51142 during light-dark and continuous-light growth. C. (2005). & Lane. 9(4). Zhao. 1-20.. Regulation of metabolic networks: Understanding metabolic complexity in the systems biology era.. X. P. 499-507. Zorin. (2007).P. et al. Shimizu. et al. P. J. I. et al. 20(5).S. G. Ort.. Methods in enzymology. Gaucher. 379–407. Jane. Turner. (2002). 1264-1277. 669-681. (2006). Fernie. (2005). C. T.G. E. F. L. Prometrics.F. Vigeolas. Improving gene annotation using peptide mass spectrometry. M. Zhu. L. C. Effects of growth condition on the structure of glycogen produced in cyanobacterium synechocystis sp. (2007). Fermentation metabolism of the unicellular cyanobacterium Cyanothece PCC 7822. D.A. 19(2). Plant Physiology. Zaslavskaia. et al. (2004). 4(7). Gene modifications and mutation mapping to study the function of photosystem II. K. J. (2008). Toepel. Vermaas. 325–331. (2002). W. M. 515-524. 36(2). 28 2.A. International Journal of Biological Macromolecules. Genome Research. et al. Optimizing the distribution of resources between enzymes of carbon metabolism can dramatically increase photosynthetic rate. A. (2009). H. Zhekisheva. Long. 69(2). Differential transcriptional analysis of the cyanobacterium cyanothece sp. 136(1). 40(5).. Tamagnini. T. Accumulation of oleic acid in haematococcus pluvialis (chlorophyceae) under nitrogen starvation or high light is correlated with that of astaxanthin esters.Sweetlove. et al. Hegemann. S. Plant Physiology. X.A. Long. 91–124. (2007).G. H. Xu. et al. Simmons. et al. Protist. (2006).R. et al. 157(2). M. Genes and development. E. M. 21(10). New Phytologist. What is the maximum efficiency with which photosynthesis can convert solar energy into biomass? Current Opinion in Biotechnology. Zehmer.. 66(1). 498-504. Eukaryotic Cell. Triacylglycerol accumulation and profiling in the model diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum (Baccilariophyceae) during starvation. Genes & development. Wu. S.R. A numerical simulation using an evolutionary algorithm.

algae can be cultivated via maintenance is likely to be superior to that in open ponds photoautotrophic or heterotrophic methods. Both types of cultivation systems must contend with require light to grow and create new biomass. important that cultivation R&D they do not receive the benefit of evaporative cooling projects are closely associated with techno-economic and so temperature must be carefully maintained. Photobioreactors can also provide a various features of both approaches). it is important to 3. reducing the amount of water that Photoautotrophic Cultivation must be processed and thus the cost of harvest (Christi. Many of these issues are being for closed photobioreactor construction are currently higher addressed through improved material usage and enhanced Exhibit 3. are unlikely to be sterilizable and may require periodic cleaning due to biofilm formation. it is premature to predict whether problems of scalability. Though photobioreactors (e. Algal Cultivation than for open ponds raceways. For photoautotrophic cultivation strategies where algae 2007).g. (TE) analysis that can evaluate the cultivation system Open ponds. higher surface to volume ratio and so can support higher volumetric cell densities. however. However. photobioreactors)..1 Comparative features of microalgal cultivation approaches ADVANTAGES CHALLENGES • Less loss of water than open ponds • Scalability problems • Superior long-term culture • Require temperature maintenance as they Closed maintenance do not have evaporative cooling Photobioreactors • Higher surface to volume ratio can • May require periodic cleaning due to support higher volumetric cell biofilm formation Photoautotrophic densities • Need maximum light exposure Cultivation • Subject to daily and seasonal changes in • Evaporative cooling maintains temperature and humidity Open Ponds temperature • Inherently difficult to maintain • Lower capital costs monocultures • Need maximum light exposure • Easier to maintain optimal conditions for production and contamination prevention • Cost and availability of suitable Heterotrophic feedstocks such as lignocellulosic sugars • Opportunity to utilize inexpensive Cultivation • Competes for feedstocks with other lignocellulosic sugars for growth biofuel technologies • Achieves high biomass concentrations 3. open gas exchange (both CO2 and O2). photobioreactors have suffered from and cyanobacteria. Photobioreactors to identify the best-suited cultivation system.1 compares the readily introduced. Algal Cultivation  29 . are subject to daily and seasonal in the context of upstream and downstream processing changes in temperature and humidity. It is. open ponds) or hybrid systems will prevail in the lose much less water than open ponds due to evaporation.1 Cultivation Pathways acknowledge the advantages and disadvantages of both photoautotrophic cultivation approaches. but long-term culture Broadly speaking. capital costs maximizing light exposure. Microalgae and Cyanobacteria Given the multiple pathways to cultivating microalgae Traditionally. therefore.g. industry. especially in terms of mixing and algal cultivation in closed (e.. both varying in where contamination and “foreign” algae are more their challenges and advantages (Exhibit 3.3.

1993. Reddy et al. using artificially upwelled water produce a variety of products at large scale. 1993) and the phycocolloid-producing seaweeds pond and photobioreactor systems have been published or Gracilaria. Land-based pond systems have also been considered for macroalgal cultivation (Friedlander. which harnesses both the overcome due to the conflicting uses of coastal zones. important seaweeds such as Undaria (Kawashima and 30 3. (Chynoweth. such as sugars... Saccharina latissima and Nereocystis luetkeana have in greenhouses are attached to substrates (usually rope been successfully co-cultured with salmonid fish species structures)... and Chile. in particular. biomass concentrations that reduces the extent and cost of 2004. 2007). Sargassum. Laminaria hyperborean. Macroalgae 1987). Kumar et al. Macroalgae can be cultivated in off-shore. efforts. 1993).. As a result.. Heterotrophic cultivation also achieves high successfully tested in the North Sea (Buck and Buchholz. Turbinaria. already widely used to Southern California.. to generate Biomass Program through several deployments of kelp new biomass. Near-shore coastal environments are already The primary challenges with this approach are the cost and being exploited by countries like China. encountered with the stability of either the structures Optimal conditions for production and contamination themselves or the stability of the attachment of kelp to prevention are often easier to maintain. located in close proximity to conversion operations Although the field is still at an early stage of development. providing high cell density algal inocula promising for growing plants from protoplasts (Dai et for production ponds (Ben-Amotz. 1995) or a series of al. and culturalists to advance the vegetative propagation disease.. have been algal growth. thus providing optimism for future the infrastructure required to grow the algae (Xu. Dipakkore et al. Modern integrated aquaculture scenario in co-culture with finfish macroalgal cultivation technology that is based on the use and mollusks. In sugars. they could compete for feedstocks and popular resistance against use of coastal regions for with other biofuel technologies. considerable scale-up from current activities. 2007). Porphyra been in practice since the 1950s. While it was determined that such cultivation presents a different set of advantages and structures would support growth of kelp. linked turbidostats or chemostats (Benson et al. et al. 2005. However. The operation of large offshore In heterotrophic cultivation. and presented (see Chapter 10). and predation. Dai et al. in hybrid al. it remains to be seen which system protoplasts is possible in both the laboratory (Dipakkore will be preferred at scale over long periods of operation. 2004. 2008). Demonstrations of successful callus formation and improvements in efficiency of water movements and plantlet regeneration have been reported in commercially pond construction costs are needed (Friedlander. 3) ease of nutrient application without dilution. and 5) possibility of farm operations of seaweeds through cell and tissue culture techniques. 2005).. modern prototypes for offshore potential to utilize inexpensive lignocellulosic sugars for growth of the kelp. both as free-standing algal farms and in an Macroalgae require unique cultivation strategies. 2) use of plants with or without holdfast Modern tools developed in the larger plant breeding structures.. and there is the the structures. 2002). appears especially open raceways. been adopted by seaweed biologists (Garcia-Reina et al. near-shore. and major technological 1991). over those in water have been listed as 1) ease of plant management. algae are grown without light seaweed farms was initially tested by the Marine and are fed a carbon source. Hypnea. 2004. or Heterotrophic Cultivation in open pond facilities. Recent studies have play a critical role as breeder/feeder systems linked to shown that Porphyra. availability of suitable feedstocks such as lignocellulosic which have viable seaweed aquaculture industries. wastes from the other species of artificially produced seed as a source of propagules has represent a nutrient supply for the macroalgae. Japan. A related approach aquaculture represent challenges that will need to be is mixotrophic cultivation. then reared to plantlet size and transplanted (Bruton et al. 2006) and field Additionally. Typically. improvement in strain selection. difficulties were challenges compared with photoautotrophic methods. Growth of plantlets regenerated from data. seeds grown al. 2004.. 2005) designs. 2006). much of the information used Gelidiella (Collantes et al. Hanisak. for these analyses is based on assumptions or proprietary Kumar et al. Because these systems rely on primary productivity the United States and Europe. photobioreactors could (Dai et al. Algal Cultivation  . Advantages of the land-based systems to coastal farms for grow-out to harvestable size. 2008. Though TE analyses for both open Tokuda. environmental regulations from other sources. This approach takes advantage of mature on growth structures in deep waters off the coast of industrial fermentation technology.. Heterotrophic as a nutrient source. Buck. 2004. In the latter. photoautotrophic and heterotrophic ability of algae. For contribution to a biofuels the micropropagation of plants is a concept that has marketplace. community are now available to seaweed biologists 4) avoidance of open sea problems such as bad weather. Dai et al. 2009).

or and take over bioreactors and open ponds? heavy metals into the biomass stream (Hoffman et al. Some members of the a mixed or natural assemblage of organisms in an attempt community will be of positive value. Wilson et al. proteins. and be necessary in open systems since seasonal variation in • Water conservation. Fungal of supplemental CO2 in large-scale algal cultivation and viral pathogens are common. viable. Boutte et al.. e. for real-time monitoring.Other options for algal cultivation are being investigated Stability of Large-Scale Cultures including the harvesting of naturally occurring marine algal Systems for large-scale production of biofuels from algae blooms.000 and several million phytoplankton 3. Wilson et al. and inexpensive.. sustainability and FISH (Marie et al. Because of the and even less is known about host resistance mechanisms. predators.. 2009). The cost-benefit analysis al. competitors. limited. One of the more into a variety of useful products. In certain cultivation systems. management.. 2006.. Continuous monitoring will management. 2009). little is known about might limit such takeovers? artificial pond ecology or pathology. 2008).. and the use of algal mat or biofilm cultivation must be developed on scales that are orders of magnitude schemes. Rittmann et al. 2008. and investigation into these areas will be important for the development of large- scale cultivation risk mitigation and remediation strategies. commercial-scale algal cultivation: selective. (2009) point out that though there may be between 40. especially in open larger than all current worldwide algal culturing facilities systems. pervasiveness of issues related to scale. or flow-cytometry-based taxonomic assays.. Tapping into existing agricultural or reservoirs of algal pathogens and predators? municipal waste streams will lower nutrient costs but could • To what extent will local “weedy” algae invade introduce unacceptable pathogens. It should be noted that. • What prevention or treatment measures Wilson et al. as well as potentially provide • Culture stability. and simple and complex carbohydrates. predators. 3.g.. 1994. The methods must be sensitive.. Chytrid fungi have also been known to cause The inherent difficulties of scaling up from laboratory the collapse of industrial algal cultivation ponds (Hoffman to commercial operations present both technical et al. but very little is known about host specificity and economic barriers to success. Nutrients. and management of competitors.2 Scale-Up Challenges species. chemical compounds. must also be managed in a way that balances is the fact that algal predators and pathogens are both productivity and pathogen sensitivity with the plasticity of pervasive and little understood (Becker. TSA- • Nutrient source scaling. 2004. 2004). 1999. and lipids still others will cause culture disruption. 2008) productivity analysis. 2006 ). it will be and require significant investment in methods for detection challenging to maintain algal monocultures on this scale. there have only been 150 formal descriptions of phycoviruses. an investment in “open source” test bed facilities for public sector Important questions concerning this threat RD&D may foster more cultivation research. Algal Cultivation  31 . This model would can scavenge and recycle nutrients or synthesize essential require a downstream biorefinery capable of processing vitamins. including worrisome components of large-scale algae cultivation CO2. Methods for rapid. 2005).. although current has yet to consider the intricacies of biological carbon understanding of their diversity and host range is very concentration mechanisms (Wang and Spalding. and pathogens is expected (Hoffman et al... Viprey et al. Others will compete for shared resources. such as those that to maximize total harvested biomass. Honda et algal physiological adaptation. automated or semi-automated biological and chemical monitoring in production settings will Four broad cultivation challenges have emerged be essential for assessing the health and compositional that are important to address for economically dynamics of algal cultures. and recycling. and pathogens. 2005. “Environmental” DNA sequence analysis can contribute to the development of PCR-based • Standardized metrics for system-level (Zhu et al. 2008. 2008. to large-scale algal cultures include: Nutrient sources and water treatment/recycling are • Are agricultural or municipal waste streams—a technically trivial and inexpensive at small scales and potentially significant source of nutrients for algal yet represent major technical and economic problems at cultivation—actually a liability because of significant commercial scales. Brussaard. Additionally. it may become necessary to understand and manage the One possible approach to contend with this is to cultivate communities that will be present. monocultures are inherently difficult to maintain combined. Cheng et al...

sustainable fuels. cultivation schemes should lead to enhancements in productivity. diverging dynamics will be important. need to be more rigorously culture prior to water recycle. vitamins. a significant investment toward basic of algal lipid contents (Gocze and Freeman. as siting requirements for efficient considered. There is justification to carry out such as sulfur. and production siting.. and genetic. Thus.. The development of studied. New approaches are split Oswald. developing an understanding of pond transportation and the volatile market for carbon credits will speciation. nitrogen nutrition has long been known to affect lipid accumulation in phytoplankton Nutrient Sources. Reed et al. Strain selection should take nutrient requirements into account. trace metals.. and ecology be a major challenge for TE feasibility studies.. 2009) and a focus especially important issue as there have been calculations on better understanding the mechanisms of biological that the world’s supply of phosphate is in danger of running CO2 concentration from ambient air (Lapointe et al. Sheehan et al. coordinated research amongst analytical System Productivity chemists. Algal Cultivation  . predators. phosphorous. derivatives of the Bodipy for both wild-type and genetically modified algae. Requirements for additional nutrients. physiologists. but vary depending upon the specific strain and algal cultivation may rarely coincide with high-volume water source chosen. biochemists. Spalding 2008). automated. molecule with higher lipophilicity or lower quantum yields whose competitiveness in the field cannot be accurately in aqueous solvent may prove to be more reliable indicators predicted. iron. Gao et al. sustainability. Phosphorous appears to be an (Rosenberg et al. fit-for-purpose fuels. phosphorous. accounting for 6-8 cents per this area is receiving new attention from the search for gallon of algal fuel in 1987 U. 1938. The synthesis of new non- genetic modifications of production strains may become toxic. competitive Magnetic Resonance-based methods for rapid lipid content production strains.. The use and availability of carbon- point sources of CO2 (see Chapter 9). 2002).. Utilization of existing and in the development of an algal biofuel industry could new knowledge related to the physiological regulation of facilitate the deployment of algal based-biofuels by lipid or carbohydrate accumulation coupled with scalable ensuring consistent. This information will productivity of potential strains. must also be R&D in both areas. and significant operating cost. productivity lost during down time. Benemann and Oswald. models and establish protocols for measurement of yields. The cost of CO2 based nutrients for heterotrophic growth will also affect the economics and sustainability of such systems.S.. and silicon (in the case of diatoms). For example. civil and mechanical engineers is needed for cultivation science and engineering will yield significant rapid progress. Shifrin and Chisholm Sustainability. Nutrient supplies for algal cultivation have a sizeable 1998). 1999). and pathogens will be a key to the success of remedial actions and for determining Better methods to detect the amount of desired fuel when decontamination and subsequent restart procedures precursor produced will be required to assess the represent the only alternative. light stress in some marine phytoplankton may also The primary focus is the major nutrients – nitrogen. These tools. and iron additions represent a long been known to increase algal growth rate. rates. result in increases in lipid content (Azachi et al. Research at the interface between basic algal biology and chemical. Along with standards. and normalization. supplemental CO2 has Nitrogen. This calculation takes into account a 50% between using algae to scrub CO2 from emission gasses rate of nutrient recycle. National and international efforts toward improvements in productivity while at the same time generating quality assurance policy standards early on lower the cost of production. 32 3. From a productivity standpoint. inexpensive chemical treatments or physiological adaptations and high-throughput monitoring. fluorescent indicators other than Nile Red necessary. 1987. 1999. 1996). 2008. and products. 2008). out (Abelson. densities. research in multi-trophic. 1994). as well as others such potential need to either discard or treat the contaminated as Near Infra Red spectroscopy. predator-prey relationships. More recent data suggest that high salt and high impact on cost. Eltgroth et al. dollars (Benemann and renewable. Fluorescent and Nuclear also inform efforts at developing robust. metabolites. and Management 1981. permeable. 2008. Douskova et al. and because of the 2005. Early detection schemes for business models are already apparent on these issues. 1996. invasive species. (Ketchum and Redfield. and adapted for rapid. etc..Furthermore.. molecular-level algal ecology will be an important component of the investment There is an immediate need to standardize productivity portfolio required for developing the potential of algae. Dynamic pond monitoring will be important are also important. For example. The frequency of contamination screening in algae have been developed and applied to events that require decontamination/restarts will be an many different types of phytoplankton with mixed results important parameter in the cost of production because of (Cooksey et al.

but One of the main advantages of using algae for their costs are still significant. Further research into the availability day (Weissman and Tillet. also become necessary to expand the limits of analysis to include recycling of nutrients from animal waste. the culture medium pose a problem for waste water energy. for example. If this blowdown water contains substantial compatibility with cultivation sites. biogas.. Note also that flue gas fed to algal cultures may particularly problematic for recycling of silicon. On the other hand. too much of a particular nutrient may prove toxic. Finding inexpensive sources of nutrients will be important. Algal Cultivation  33 . aquifers and seawater. Though the water used to initially fill the pond can be saline.g. From a to fossil fuel prices. or urea. by preventing eutrophication. but it may also be desirable to use nutrient levels are vitally important factors for cultivation limitation (e. For example.. or silicon) as a economics. Although economics dictate that the bulk of nutrient sources and recycling scenarios water derived from the harvesting step must be returned • Studies to explore the mechanisms of nutrient to the cultivation system (where remaining nutrients recycling. evaporative losses can exceed 0. however. Reagent grade sources of nutrients could make the price Conservation. and the very large required energy sustainability perspective. it may NOx and SOx (Doucha. which these nutrients are re-mobilized and made available which may or may not be compensated for by the “free” for algal growth are not well understood. nitrogen. unused nutrients in • TE and life cycle analysis to understand the cost. Currently.Nitrogen is typically supplied in one of three forms: ammonia. real economic or sustainability problems or because of Utilizing agricultural run-off also poses economic benefits loss of public support due to perceived problems. a certain amount • Geographic Information System (GIS) analyses of of “blowdown” water must be removed to prevent salt wastewater resources to understand availability. Because synthetic nitrogen fixation processes contains no nitrogen. then most of the nutrients will remain in the such a scheme will certainly have some impact on overall digestor sludge and can be returned to the growth system productivity levels as photosynthetic energy could (Benemann and Oswald. 20 metropolitan areas with high land prices and limited land cm deep open pond will require 530. Also. phosphorous. However. At the same time. and sustainability implications of various discharge. 2005. nutrient recycle may prove inputs should be accounted for in life cycle analyses. tend to be near enormous. If the biomass residues are. such as saline water from waste streams is a very attractive alternative. brackish. availability. algae are used in some wastewater treatment facilities water management poses some of the largest issues because of their ability to provide oxygen for the bacterial for algal biofuels. and potential nitrogen and phosphorous. from anaerobic digestion sludges can feed subsequent algal growth). phosphorous. produced 3. nitrate. and sustainability issues. for example. gallons per day from the 1 ha pond.000 and compatibility of wastewater resources is warranted. With large cultivation systems. agricultural. water demands will be Wastewater treatment facilities. The final fuel product from algal oil biology. these nutrients utilize fossil fuels (particularly natural gas). and it is not practical to transport wastewater In desert areas. Douskova et al. disposal will become a problem impact of such sources on algal biofuels production due to concerns of eutrophication of surface waters. e. A potential problem with this approach however is the impact on facility siting.. a hypothetical 1 hectare (ha). or industrial unsuitable for land crops. Therefore. 1998). and Recycling of a gallon of algal fuel cost-prohibitive. or iron. cyanobacteria as a way to provide nitrogen biologically. to be more valuable than using the spent biomass for It is possible to consider the use of nitrogen-fixing products such as animal feed. productivity. which is provide some of the nitrogen and sulfur needed from a component of the diatom cell walls. utilizing the biofuels production is their ability to thrive in water nutrient content of municipal. costs are tied end up primarily in the spent algal biomass.5 cm per over long distances. Agricultural- or commodity-grade nutrients are more applicable. 2009). 1989). which is a loss of 13. The ideal form of nitrogen Another approach to reduce nutrient costs is to pursue is a function of relative costs and the specific strain’s a diligent recycle. If not addressed adequately. 1996).000 gallons to fill.” either because of and phosphorous into biomass for water clean-up. Careful control of nutrient levels is also critical. Water Management.. This may be nitrogen. buildup.g. Limitation of a key nutrient will have serious impacts on biomass Nutrient sourcing and the control of nutrient productivity. In the future. treated by anaerobic digestion to produce perhaps in co-culture with eukaryotic algae. water breakdown of organic materials and to sequester nitrogen can easily become a “show-stopper. means to induce oil accumulation in the cells (Sheehan Important research areas therefore include: et al. The processes through be diverted from carbon fixation to nitrogen fixation.

flocculants. Furthermore. or other low. toxins (including heavy cooling. including saline Conservation of water can be addressed to some extent aquifers. meaning that half movement (pumping costs) the culture volume must be processed daily. The blowdown water exiting the process will also most likely require treatment. accumulated salts. groundwater.. however. etc. and continually making up the volume with affect biodiversity of neighboring ecosystems or result low-quality water will concentrate salts. chemical flocculants used in harvesting. moving around such large volumes of water is very energy-intensive and can impose a significant cost. This is an enormous amount of water (260. from flue gas). To contain costs. This could include reducing part of the analysis that goes into siting an algal facility evaporative cooling loads through such strategies will be to analyze the “pan evaporation” rates at specific as selecting thermotolerant strains of algae sites in conjunction with water cost and availability.000 gallons per day in the 1 ha example above). makeup water/recycle. The added cost of such systems must down saline aquifers. research in the following areas is warranted: in the culture and the salinity of the replacement water.) of the recycled stream could be cost-prohibitive. desalination.. municipal wastewater. activated charcoal filtration. while effectively Water recycling is essential. • Studying water recycle and methods to maximize recycle (and minimize blowdown). or seawater) may be suitable as is. Note however • Analysis and definition of the regulatory landscape that evaporation plays a critical role in temperature surrounding discharge of water containing various maintenance under hot conditions through evaporative levels of salt.water from oil wells. and water double their biomass on a daily basis. and treatment (e. heavy metals (e. it is desirable to recycle most of that water back to the culture. and residual live algal cells. accumulated toxics. or other remediation before use. Incoming water (surface water. residual nitrogen and phosphorous fertilizer. Closed systems that spray water on the surfaces metals). An advantage of sites that may have lower pan evaporation rates closed photobioreactors over open ponds is a reduced • Understanding the long-term effects of drawing rate of evaporation. flocculants. Disposal of the spent water.g. Surface disposal and reinjection into wells may be an option as regulated 34 3. Algal Cultivation  . or may require decontamination. This can be prevented by adding Sterilization of blowdown water. Some actively growing algal cultures can treatment. water. and live cells or employ cooling towers to keep cultures cool will lose some if not possibly all of the water savings of such • Developing cultivation systems with minimal systems under these conditions (Flickinger. which could contain salts. by the Environmental Protection Agency and already quality water stream. However. materials in the culture.1999). • Investigating ways to reduce the cost of water and location. by disposing of a portion of the pond volume each day as “blowdown.” The amount of blowdown required for Because of the importance of issues surrounding the use salinity control is dependent upon the acceptable salt level of water. or biological inhibitors produced by the strains themselves could impair growth if recycled to the culture. including the geology of these be balanced against the cost savings and sustainability aquifers and associations with freshwater systems analysis for water usage for a given location. wastewater. but the amount that can managing the accumulation of salt and other inhibitors be recycled depends on the algal strain. and other in the dissemination of genetically modified organisms. • GIS analysis of water resources.g. toxins. process. would be a fresh water—a costly and often unsustainable option—or very costly and energy-intensive proposition. but live cells could adversely is fresh water. the water being lost to evaporation practiced by the oil industry. Treatment may be essential for water entering and exiting the process. disinfection. A critical water consumption. and their proximity to utilizable cultivation through facility design and siting. could pose a serious problem.

ed. B. P. {Bibliography}Douskova. B. Guckert. 7. Journal of Applied Physiology. Z. Aquaculture. Final Report to US DOE NETL. Cambridge. Benson. Buchholz. 6(6). V. C. R. Hino (2004). 2005. Wu. 395–419. 17. Journal of Applied Phycology. Watwood and G. Biotechnology Advances. 139-145. Shen. K. D. (2004). 16. P. Z. Aflalo. K. Journal of Microbiological Method. P. James and S. J. 221:181-194. Applied Microbiology and Biotechnology. Maeda and A. C. Israeli R & D activities in seaweed cultivation. Fisher. (2006). Genetic breeding and seedling raising experiments with porphyra protoplasts. 542-550.. 331-337. Viral control of phytoplankton populations: A review. Hydrobiologia. (2008). D.. P. 203-213. V. M. 36(2). J.. Cooksey. M. 1000-1009. Chynoweth. Salt induction of fatty acid elongase and membrane lipid modifications in the extreme halotolerant alga Dunaliella salina. 179-185. (2007). Zamir. 3. Zarka. Hoffman. J. 41(5). Aoki. Buck. potential and speculative applications of seaweed cellular biotechnology-some specific comments on gelidium. Callis. S. The Journal of Applied Phycology. Micropropagation by explants of Gracilaria. Biodiesel from microalgae. K. 125-138.References Azachi. Zhang. Goldshlag. Actual. B. Xiong. 1320-1329. Sadka. M. (1994). 331-343. Hanisak. Livansky. Benemann. Plant Physiology. C. R. M. K. Buchholz. Rhodophyta) in laboratory culture. and Z. Machat. Freeman. The Journal of Eukaryotic Microbiology. Dipakkore. (1987). Factors underlying the variability of lipid droplet fluorescence in MA-10 lydig tumor cells. B. I. McLachlan and Oliveira. 191-218.. Cambridge University Press. Cheng. R. M. S. Boussiba. Q. Melo. Production and cellular localization of neutral long-chain lipids in the haptophyte algae Isochrysis galbana and Emiliania huxleyi. Competition between the rotifer brachionus rotundiformis and the ciliate euplotes vannus fed on two different algae. Zhou. Bao. and A. X. Candia. Gocze. Boutte. (2008). C. Simultaneous flue gas bioremediation and reduction of microalgal biomass production costs. Encyclopedia of bioprocess technology: Fermentation. Elsevier.. Robledo. Brussaard. Eltgroth. (2008). Sosa.. 56. 250.. Chilensis Bird. Y. J. 82(1). Novak. 512. Grubisic. Collantes. 65-68. 241(1-4). H. Jha. G. Y. University of Florida. 112. C. E. 355-368 Buck. Han. Balthasart and A. Flickinger MC. and P. D. 15-28.. Review of Biomethane from Marine Biomass. (2004). Gomezpinchetti. Rapid quantitation of lipid in microalgae by time-domain nuclear magnetic resonance. Chisti. Wilmotte. D. 51. 2002. Gokhman and A. A. Journal of Microbiological Methods. P. Algal Cultivation  35 . Wiley. Aquaculture. Amsterdam. Y. W. and B. (1994). (1996). (1999). T. Ben-Amotz. R. Rusch. S. Umysova. (2004). X. Zhang. L. The offshore-ring: A new system design for the open ocean aquaculture of macroalgae. 17. 198-211. M. A. Gutman. Microalgae: Biotechnology and Microbiology. Gutierrez-Wing and K. J. A. 16. S. Response of offshore cultivated laminaria saccharina to hydrodynamic forcing in the north sea. C. L. E. R. Williams and P. 75(3). G. Y. W.. Hydrobiologia. The Journal of Applied Phycology. Liu. A. Wolfe. F. S. (2004). Zachleder and M. J. Vitova (2009). H. Israel Journal of Plant Sciences. 333-345. M. Y. M. (2005). in Seaweed Cultivation for Renewable Resources. and C.. J. 127-131. T. 674-691. Cultivation of gracilaria and other marcoalgage in Florida for energy consumption. Production and seeding of protoplasts of porphyra okhaensis (Bangiales. M. T. J. and C. and P. Reddy. L. Testing of primers for the study of cyanobacterial molecular diversity by DGGE.. Aquaculture Engineering. 1-207. 151-158. Yuan and Q. P. Fluorometric-determination of the neutral lipid- content of microalgal cells using nile red. W. Benson. (1993). Cytometry. S.. Q. and L. Doucha. Yang. M. and D. 65(3). 111. Bird. Aquaculture Engineering. 129(3). 294-306. (2005). parasitic on the green alga Haermatococcus. H. The Journal of Applied Phycology. Garcia-Reina. Mycological Research. Q. (2007). A. Q. W. Systems and economic analysis of microalgae ponds for conversion of CO2 to biomass. Gao. New mode of Dunaliella biotechnology: Two phase growth for beta- carotene production.. Drew SW. The development of a mechanistic model to investigate the impacts of the light dynamics on algal productivity in a hydraulically integrated serial turbidostat algal reactor. Bao. Dai. Friedlander. M. A. (2002). D. J. H. Becker. S. D. C. and W. Isolation and characterization of a novel chytrid species (phylum Blastocladiomycota). Dai... 437-440. Oswald. 25. L. (1991). R. Journal of Microbiological Methods. biocatalysis and bioseparation. M.. B. Seedling production using enzymatically isolated thallus cells and its application in porphyra cultivation. (1995). (1987). 70-81. I.

FEMS Microbiology Ecology. Dunahay. Callus induction and thallus regeneration from callus of phycocolloid yielding seaweeds from the Indian coast. Lapointe. Phytoplankton lipids-Interspecific differences snd effects of nitrate. and B. M. phaeophyta). 261.. Mapping of picoeucaryotes in marine ecosystems with quantitative PCR of the 18S rRNA gene. An external delta-carbonic anhydrase in a free-living marine dinoflagellate may circumvent diffusion-limited carbon acquisition. San Diego.. Thiruppathi. 253-268. Yokochi. 15-25. Dipakkore. Marie and D. E. Petty. Biological Bulletin. Wide genetic diversity of picoplanktonic green algae (chloroplastida) in the Mediterranean sea uncovered by a phylum-biased PCR approach. Wilson. 1427-1436. J. L. rhodophyta).. (1993). G. Department of Energy’s aquatic species program-Biodiesel from algae. Reed. C. silicate and light-Dark cycles. D. Phycologia. J. A method for maintaining a continuous supply of marine diatoms by culture. The Journal of Applied Phycology. Redfield. L. K. 6. (1998). Vaulot. Nakagiri. W.. Not. Reddy. Wilkinson and M. and post-genomic study of microbial communities involved in bioenergy. J.S.. Academic Press. F. (2004). Eswaran. (1999). (1989).. A. 10110-10115. P. Higashihara. (2006). K. Weissman. 52(1). 328: 1-42. Algal Culturing Techniques. H. Proceedings of the National Academy of Sciences. A. Y. Sheehan. 43. X. (2008). and Wu. Graham and R. 430-436. Journal of Phycology. Retrieved June 29. Bot. 604-612. Berlin. J. A look back at the U. Ganesan.. S. Benemann and P. and A. (2007). C. Tokuda. G. 499-507.: Viprey. Q. Van Etten and M. W. 75(1). N. T. Hydrobiologia. from www.. Rosenberg. N. (2008). Environmental Microbiology. Marie. D. Coury. Plant Physiology. K. 385-389. Regeneration from callus of undaria pinnatifida (Harvey) Suringar (laminariales.. 10(7). Krajmalnik-Brown and R. (2005). M. Phytoplankton cell counting by flow cytometry. U. S. Massana. Springer-Verlag Berlin. Raghukumar. (1981). Brzezinski. T. D. Vaulot. Nature Reviews Microbiology. Schaumann and T. 374-384. Ketchum. 36 3.. T. J.. K. R. An inorganic carbon transport system responsible for acclimation specific to air levels of CO2 in Chlamydomonas reinhardtii. M. Guillou.. Morse. Halden. Vaulot.nrel. Pre-genomic. (2005). C. Rittman.. W. C. 46(6). R. B. Roessler. J. MacKenzie and D. Lesser Known Large dsDNA Viruses. S. Ferreol and D. Microalgal carbon-dioxide-concentrating mechanisms: Chlamydomonas inorganic carbon transporters. D. M. Chisholm. Jha. Betenbaugh.pdf. Marine Biology. CA. Shifrin. R. (2009). J. Oyler. Exp. 103. 79-92. N. and R. Jha. Martin H. Journal of Eukaryotic Microbiology. (2006). Reddy. 147(3). (2007). Spalding. B. 596-602. A. R. 126. 165-169. Nakahara. G. Xu. 2007. and S. 637-647. M. Algal Cultivation  . B. S. Tissue culture and regeneration of thallus from callus of gelidiella acerosa (gelidiales. Journal of Biotechnology.Honda. T. L. Wang. J. R. Neutral lipids in macroalgal spores and their role in swimming. L. and M. Miao. (1999).. 1804-1822.. D. 133(4). A. H. (1938). High quality biodiesel production from a microalga chlorella protothecoides by heterotrophic growth in fermenters. (2008). Tillet. M. M. C. and H. Y. A green light for engineered algae: Redirecting metabolism to fuel a biotechnology revolution. Spalding. The Phycodnaviridae: The story of how tiny giants rule the world. Allen. S. and H. (2008). F. H. Rao. Kawashima. A. Kumar. Design and operation of an outdoor microalgae test facility. D. H. Molecular phylogeny of labyrinthulids and thraustochytrids based on the sequencing of 18S ribosomal RNA gene. Current Opinion in Biotechnology 19(5). 737-744. genomic. 17(4). R. 19. Zhu. Kumar. Simon and D. Anderson .

The final slurry concentration will depend on the extraction methods employed and will Optimizing flocculation methods. type. and systems to liquid transportation fuels requires processing autoflocculation in the form of photosynthetically driven steps such as harvesting (Dodd and Anderson.. Their small sizes. gm water) must be concentrated to slurries containing Mollah et al. Flocculation and Dissolved Air Flotation A number of different forms of forced flocculation have been employed. 2004. energy costs climb steeply. Chemical additives that bind algae or Flocculation and Dissolved Air Flotation (DAF) was otherwise affect the physiochemical interaction between established for sewage treatment and later studied in algae are known to promote flocculation (Lee et al. Bare et al.. Likewise. Alum. at least 1% algae given the known processing strategies (Borowitzka. 2006. Harvesting • The environmental impact of flocculant or pH Flocculation and Sedimentation manipulation in released water effluent. 1991. may analysis of engineering designs. 2005. 2001)... Culture manipulation techniques. Final will very likely depend on strain selection. algae harvesting (Sim et al. combined with RD&D to be useful for promoting flocculation.02 . 1988). being considered. 1975. Phochinda and White. Flocculation is used to increase the additives that have been studied.. and on empirical of transportation. and then air is bubbled 4. Bioflocculation where algae are co- dewatering. 1998. salts. Flocculation leading to sedimentation occurs • Optimized sedimentation tank designs with integration naturally in many older cultures. Botes and Vanvuuren. Future research in develop specific processing technologies to support those flocculation chemistry must take into account the following: designs and a fundamental understanding of how algal biology can impact harvesting and dewatering strategies. and other man-made fibers are some chemical Lincoln. additives. water quality. These energy-intensive processes are has also been considered (Lavoie and Delanoue. 1983). and extraction of fuel precursors (e. Poleman and Pauw. 1997). well-managed high growth rate cultures due to their • Bioflocculation. A determinations in particular processes. polyacrylamide polymers. CO2 depletion for pH control has been studied (Sukenik Butterfi and Jones. Pan et al. Knuckey et al.. It is possible to feasible algae-to-fuel strategy must. 1984). consider the imagine selecting/designing strains to aggregate on cue energy costs and siting issues associated with harvesting or designed with a particular flocculant interaction in and dewatering. Downstream Processing: Harvesting and Dewatering  37 . the mechanism slurry concentration also impacts plant location because of algae-flocculant interactions. Downstream Processing: Harvesting and Dewatering Conversion of algae in ponds. This facilitates the transport of electrocoagulation must be scaled-up with cost and cells to the photoactive zone through pond or bioreactor energy analysis. only now being recognized as critically important. • Chemical flocculant recovery techniques are required Processing technologies depend on the algal feedstocks to minimize cost and control water effluent purity. therefore. is required to promote sedimentation at harvest. surfactants.0. and recycling issues. 1971).1 Approaches for Microalgae further downstream extraction and fuel conversion processes must be understood and controlled.g. mixtures.07% algae (~ 1 gm algae/5000 the advantages of no added chemicals (Chen. 1969. electroflocculation and electrocoagulation offer with as low as 0. Cultures Finally. 1988. and off-shore pH with and without additives is also effective. some into further downstream dewatering techniques. Processes that pertain to unicellular algae are quite different from the approaches applicable • The effect of residual flocculant or pH manipulation to macroalgae... and Shelaf. Addressing these issues requires careful mind. McGarry and Tongkasa.4. 2004. Koopman and chitosan. 2003. 1993. and fuel Microalgae and cyanobacteria remain in suspension in conversion and use must be considered. cellulose. Manipulating suspension size of the algae aggregates. and chemistry to maximize algae recovery of dry biomass increases. water form of forced flocculation usually involving chemical recycling and flocculate recovery are required. Edzwald. As the desired percentage concentrations. and small size (~1 to 30 µm). bioreactors. 1977. in recycled water on culture health and stability and lipid production must be understood and controlled. make harvesting more difficult. impact the required energy input. electroflocculation. therefore. 1987). the presence of flocculant in 4. circulation. lime. In managed cultures. Kwak et al. lipids cultured with another organism that promotes sedimentation and carbohydrates). however.

but will require extra space and • Filter material also influences filtration and recovery considerable time.. Significant cost and engineered design for further downstream processing.2 Approaches for Macroalgae blinding are also issues. in low-humidity climates.through the suspension causing the algal clusters Centrifugation to float to the surface. and flocculant reclamation) are also investments. leads to blinding. • The filter pore size is critically important as it is defined by the size of the algae species and algae Drying aggregation rate. and have the ability to remove the algae later. Materials can be used that optimize filtration energy are also possible. 2009). 2001). where fuel precursors very expensive. Solutions involving either solar or wind efficiency. such mechanized harvesters require boats should consider minimal or no washing requirements. Centrifugation for sedimentation (e. recovery is largely dependent on current level of centrifugation technology makes this bubble size and distribution through the suspension. Application of mechanical 38 4. approach cost-prohibitive for most of the envisioned DAF facilities require optimized integration with any large-scale algae biorefineries. 2008). and mechanized harvesting methods. and reduction of filtering rates. so leads to re-dilution of the product. • An important step is recovering the algal biomass from which can involve mowing with rotating blades. the costs climb steeply with incremental becomes important as a distribution of microorganism temperature and/or time increases. 1995. Filtration designs Invariably. water and technologies must consider large initial capital equipment algae purity. 1996). Saidam and Butler. manipulation of Filtration is conceptually simple but potentially electric fields. The flocculant efficiency. 2003).. Manual harvesting is common for both cultivated and natural systems. design.. but are not limited to. shrimp and further understanding of several issues: tilapia) grown with algae (Johnson and Wen. Downstream Processing: Harvesting and Dewatering  . For instance. acoustic focusing to concentrate algae at nodes. Washing the filter is one practice. or ships for operation.. however. separations and has been investigated in algal harvesting (Molina et al. Power costs will certainly influence derived from cultivated sources (Roesijadi et al. Filtration Solid/liquid filtration technologies are well studied. filter materials with controlled hydrophobicity and/or algae affinity can be developed. the blocking of filter dryers and other oven-type dryers have been used. • Filtration design is an important variable with both Harvesting static and dynamic filtering operations. and can be optimized through are harvested from higher organisms (e. but doing or dredging with cutters. and high throughput encountered in flocculation and DAF. These include. the Microalgae and cyanobacteria present unique filtration use of organisms growing on immobilized substrates challenges because most strains considered for energy where the amount of initial water is controlled and feedstocks have cell diameters less than 10 µm. microalgae. of the roughly 1. In addition to processing of large quantities of water and algae.. pores. 2002..g. methane drum size. have also been developed. energy savings must be realized before any widespread implementation of this approach can be carried out. Decreasing pore Because drying generally requires heat. Other Harvesting Techniques and filtration without prior flocculation can be used A number of other techniques at various stages of to harvest and dewater algae (Ferguson et al. and bioharvesting. Culture purity However. about 90% is (Oswald. Modern seaweed harvesting vessels can be equipped with pumps to direct harvested seaweeds directly into nets or other containment structures (Ugarte and Sharp. Moving filters Currently. The algae-rich top layer is Centrifugation is widely used in industrial suspension scraped off to a slurry tank for further processing. Small algae pass through larger Drying is required to achieve high biomass concentrations. R&D have been proposed to harvest and dewater Downing et al.6 million dry metric tons have been used in drum and cylinder press designs of total seaweed harvested worldwide. pores decreasing filter efficiency. the growth substrate can be easily removed. 1991). Air-drying is possible size will affect filtration efficiency and blinding rates. operating costs. The efficiency is dependent on All of the issues arising from the use of flocculants the selected species (as related to size). Durability and 4.g. floc optimization. suction. the filter.

as opposed to attached to the bottom culture. adhering epifauna and other forms of Ultimately. The exact nature of such mechanization will obviously depend on the form Macroalgae have less of a demand for dewatering as of cultivation and type of algae being cultured. with higher a biofuels feedstock in Ireland (Bruton et al. strict regulations 80 .. 2009). The establishment of large offshore seaweeds may alleviate 4. To manage seaweed harvests.85% water content (Peterson et al. may be amenable to mowing.harvesters in European seaweed operations have been Milling is used to reduce seaweeds to particle sizes that are described in a recent feasibility analysis for seaweeds as \more efficiently processed. 2008). and low growing attached forms such as and reduce weight and associated transportation costs if Gracilaria will require different approaches compatible with algae are to be transported from sites of harvest to distant their growth characteristics. Downstream Processing: Harvesting and Dewatering  39 .. Further.g. 2004). maintenance. Smaller particles. could be cultivated in may be more important as a method to increase shelf-life floating pens.. and the amount of debris method. culture and end-use. with needs is required with consideration of capital equipment the degree of screening dependent on the mode of investments. 1989). 2009). Anaerobic digestion... about 20 . be harvested and the intervals between harvests to allow growth and recovery of biomass (Ugarte and Sharp. litter. Hydrothermal conversions are suited for environmental consequences of harvesting natural wet biomass and become efficient at 15 . 2001). given the issues related to sustainability and potential environmental importance as well as current cost and achievable consequences will need to be carefully evaluated. and depreciation. operations. although breakthroughs are still needed in each. The concept of large off-shore macroalgae farms and fermentation for alcohols. sand. stones. In require technology appropriate for that mode of culture.20% solids or populations of seaweed near-shore. For part of the pretreatment process. surface area to volume ratios. and hydrothermal liquefaction associated biorefineries has from the outset included for bio-oils. This will likely be a significant fraction of the will likely have less impact in procedures that can total energy cost of any algae-to-fuel process and a utilize whole seaweeds (Bruton et al.. This has important implications for plant design to answer simple • Dewatering questions like “What percentage of the total plant energy requirements or what percentage of that made available by Seaweeds immediately following harvest can have algae must be directed toward harvesting and dewatering?”. laws have approaches 90% may be necessary. attached forms that tend to stand upright. Dewatering seaweeds such as Sargassum spp.. example. anaerobic digestion and fermentation. new strategies should be developed to combine and integrate Preprocessing these processes in order to take an algae culture and The general preprocessing requirements for macroalgal convert it into slurry of a specific concentration. final algae concentration needed for the chosen extraction will likely have less debris. a unit operations analysis of energy input debris that should be removed before further processing. and hydrothermal liquefaction have either as Macrocystis. significant fraction of the total amount of energy available 4. will have higher reaction efficiency during anaerobic digestion for biogas. for a range of dry weight content based on extraction Screening for debris is considered mandatory. e. 2009). opportunities for products apart from fuels.3 Systems Engineering pressure from near-shore environments and create market While specific process technologies have been studied. biomass prior to extraction or direct conversion have This has yet to be demonstrated on a commercially been categorized as follows (Bruton et al. In forms such as Laminaria processing plants (Bruton et al. influence. which is beneficial for transportation and other Similarly. the exact ratio of water stipulated the percentages of harvestable stock allowed to to solids for marine biomass remains to be determined. mechanized harvesting techniques. 2009): relevant scale and remains a significant challenge. Although have been put in place in some countries (Pringle and some dewatering of seaweeds whose water content Tseng. by washing A critical gap is the energy requirements of these processes • Milling are not only largely unknown but unbounded.30% water content is noted to have a stabilizing harvesting may require retrieval and transport to shore. Dewatering to grown on off-shore rings (Buck and Buchholz. 2009). Algae that are grown in suspension The cost of harvesting and dewatering will depend on the culture. cultivation in land-based pond systems will processes requiring further drying (Bruton et al. shredded or milled macroalgal biomass can go directly into either reactions As a result of growing concern about the potential or extractions. such fermentation. scale of harvesting and dewatering. Floating a high tolerance or requirement for water. • Removal of foreign objects and debris.

(1977). 133. 300.M. D. and algae with much higher energy content than available drying as a function of the volume percentage of algae today. Preliminary Look at Energy Balance In spite of gaps in data precluding more detailed The energy content of most algae cells is of the order analyses.B. Jones. Micro-Algal Biotechnology. Stanley. Applied Microbiology and Biotechnology. R. Electrochemical technologies in wastewater treatment. B. Removal of clay and blue-green algae particles through zeta potential and particle size distribution in the dissolved air flotation process. Dissolved air flotation for the removal of algae and inorganic turbidity on a large-scale. Water Supply: IWSA/IAWPRC Joint Specialized Conference on Coagulation Flocculation. Logsdon. Borowitzka.J. Rasmussen.B. Robert. C. R.. It is possible to estimate the energy requirements in watt. 85(3). 31(3-4). J. Dodd. 5(4). 631-635.C. Algae. 67. Aquaculture. A.54 watt-hours/gram. T. Large-scale algae culture systems. Koopman.J. Kwon. 40 4. Separation and Purification Technology. H. W.J.. based on the latent heat References Bare. of each in algae are considered (Illman et al. Buchholz. J. et al. C. Botes. 113. and M. Lincoln. 27(10). D. Zubieta. (2009). and dissolved air flotation. C.P. 95. Water Science and Technology.. 9(3). (1988). (2004).R. Increase in chlorella strains calorific values when grown in low nitrogen media. Brown. J.. Bracco.. Harvesting of algae grown in agricultural wastewaters. Water Science and Technology. 357-394. de-watering. (2005). L. Biotechnology and Bioengineering. B.K. Possible approaches may include developing strains of hours/gram of algae for harvesting. B. Sustainable Energy Ireland. Downstream Processing: Harvesting and Dewatering  . I. (1987). (1975). M. Low cost reclamation using the advanced integrated wastewater pond systems technology and reverse osmosis. Lerat.X. Water Science and Technology: Water Supply.F. Y. H. Lundquist.M. E. Vanvuuren. Production of microalgal concentrations by flocculation and their assessment as aquaculture feeds. and proteins and the typical percentage integrated systems in order to overcome this challenge. H. (1995). Transactions-American Geophysical Union. F.A. Butterfi. Borowitzka.F Frampton. Algae removal using dissolved air flotation. flocculation and sedimentation and the belt filter press are expected to be minimal.. Autoflotation harvesting of algae from high-rate pond effluents. S. Johnson. Water Pollution Control Federation Research Journal. algal biofuel production schemes at scale will of 5 watt-hours/gram if the energy content of lipids. Jones. Kwak. Response of offshore cultivated laminaria saccharina to hydrodynamic forcing in the north sea. E. (27). 45(1). & Wen. Delanoue. G. J. (1991). J.R.S. Dublin. N. 612. S. M. Illman. Comparison of dissolved air flotation and direct filtration. (2010). Sedimentation and Flotation. E. The energy requirements for energy intensity of harvesting and drying algae. Integrated high-rate pond algae harvesting system. Kim. energy example shown below provides some food for thought balance can become an issue in systems that propose regarding harvesting and dewatering technologies. Scragg.. 852-859. Enzyme and Microbial Technology. 231. 35(3). (38). 117. along with innovative solutions to lower the in harvested biomass. A. 11. J..H. Jung. Ferguson. 713. 9(1). (2005)..J. 525-534. M. Z.L. 250.R.. Won. G. (2002). Shales. Curley. (2006). to take algal biomass and concentrate / dry it to enable downstream processing and extraction because of the high volumes of water that must be evaporated away. Middlebrooks.from algae. Bruton. Agricultural Wastes.B. Progress in Water Technology. Filtration. Cambridge University Press. Ferguson. Ireland Buck. Lavoie. 30 (7). Knuckey. likely need to implement innovative technologies and carbohydrates. Anderson. W.J. 6(1). (1993). 2000). Harvesting of scenedesmus-obliquus in wastewaters-auto-flocculation or bioflocculation.J. A quick and preliminary energy balance of vaporization of water at 0.. Lyons. A review of the potential of marine algae as a source of biofuel in Ireland. Chen. B.B. However.. 674-691. Water Science and Technology. 153. Edzwald. M.H. Downing. Green. (1983). 47(1). Ku. Oswald. coagulants. (1969). V. (2000).Y. Development of an attached microalgal growth system for biofuel production. D. T. and C. Aquacultural Engineering. bubbles. 50(11).

Poelman. Techno-economic feasibility analysis of offshore seaweed farming for bioenergy and biobased products. (1). Energy & Environmental Science. 491–515. Algal autoflocculation-verification and proposed mechanism. (2001). C. (1988). present and future perspectives of electrocoagulation.J. R. D. A. E. Science and Technology.. F.. Letters in Applied Microbiology. (19). et al.. 87. Oh. Kim. Cah. J. 1-7. (1971). 14. E-H Belarbi. Colloids and Surfaces A-Physicochemical and Engineering Aspects.R.B. Recovery of microalgal biomass and metabolites: process options and economics. M. The Journal of Applied Phycology.. (2008). (2001). and J. (1984).. 32-65. Downstream Processing: Harvesting and Dewatering  41 . M. Goh. Journal of Hazardous Materials. Lachance.G. Y.J. M. Phoochinda. Copping. Effects of harvesting method and growth stage on the flocculation of the green alga botryococcus braunii. 824. A. W. H. Pauw. Froling.S. A new approach to seaweed management in Eastern Canada: The case of ascophyllum nodosum. Battelle Pacific Northwest Division Report Number PNWD-3931.Lee. Water Pollution Control Federation Research Journal . J. S. Robles Medina.. R... C. (2004). Huang. (2003). 150(1-3). J. J. 142. A. D. A. 1987. W. C. Forster. Chuang. G. R.Y. Benemann. Environmental Technology. and Recycling. Butler. K. Sim.. Pan. (1998).. dissolved air flotation and drum filtration techniques for harvesting sewage-grown algae. H. and J. Biol. Water reclamation and algae harvesting. (27). 327. Kwon. Tester.M. Y. Biotechnology and Bioengineering. A. Resources. Vogel. D. Potential of electrolytic flocculation for recovery of micro- algae. S. China. White.. (1989). Removal of algae using froth flotation. Tongkasa. Yoon. 4. Sharp. E. C.. Overview of a workshop on production and utilization of commercial seaweeds . 199. M. 16(1).. P. Pringle. 63-70. 51.Qingdao. B. Tseng. T. Introduction to advanced integrated wastewater ponding systems.W. Comparison of centrifugation.C. Huesemann. Sukenik. Becker. Roesijadi. 1.. M. Peterson. G. G. McGarry. M. Mar 42. (1997).P. Shelaf. A. Algae removal by horizontal flow rock filters. (26). B114. Water. (20).S. Conservation. Biomass.C. W. Molina Grima E. N. Thermochemical biofuel production in hydrothermal media: A review of sub.A. Chisti. E. Dewatering characteristics of algae-containing alum sludge. Acién Fernández. 43(5).G. 83-89. Journal of Biotechnology Advances. (2008). Saidam. Ugarte. 24(5). Wu. (1996). 185. 1. Antal. Mollah. (2003).P. G. (1991). F. Fundamentals.and supercritical water technologies. 24(1). Oswald.D. Advances in Slow Sand and Alternative Biological Filtration.

osmotic shock. or for the sonication for cell disruption. using solvents to “milk” algal recovery/removal of high-value products. Extraction of Products from Algae   . and enzyme reactions (Mata macroalgae (Aresta.and laboratory-scale procedures. largely in the realm of laboratory-scale processes that serve analytical rather than biofuel production goals. application of organic solvents. Lipid Extraction sonication. As such the development of any extraction process as single cells suspended in water at concentrations below must also account for the fact that the tissue structure 1% solids. Mechanical disruption can include cell homogenizers. Additionally. shown to improve methane gas production and biosolids reduction in sludge digestion (Rittman et al. different logistical challenges arise from access. pretreatment of (Zhang et al. 2008. extraction bypass schemes that attempt to engineer algal systems techniques employed must be efficient and effective. Recent work on extraction of lipids from three different 5. and extraction algal biofuels as competitive bulk commodity.. Extraction of Products from Algae While relatively limited volumes of bioproducts are are attempting to bypass this step—the dynamics of currently produced from algal feedstocks. 2003). 2005) et al. Extraction depends on identifying the particular biological Mechanical Disruption (i. Non-mechanical methods include biofuel development. extraction techniques require concentrated substrates. Dibenedetto. 2008). base.. ultrasounds. but these techniques often rely increase efficiencies of vegetable lipid and oil extraction on freeze dried.e. Virot knowledge exists for the separation of plant biomass et al. 2010)... For waste treatment. Cell Rupture) component for extraction. some systems 42 5. pulverized biomass. microalgae and cyanobacteria may be cultivated lipid. While macroalgae are more like traditional and cell walls may present formidable barriers to solvent feedstocks. algal biomass extraction in aqueous phase systems serves as a starting suffers from a lack of well-defined and demonstrated place for industrial-scale extraction operations.. This generally requires that the native structure their offshore production and harvesting. 2010). any extracting solvent must be able to (1) fuel conversion process. extraction of lipids from microalgae (Lee et al. For this reason. Lipid extraction includes the following approaches: Existing extraction techniques are mainly suitable for solvent-based extraction relying on microwaves and or analytical.1 Current Practices for types of oleaginous microalgae compared bead beating. The use of microwaves to disrupt cells and have been carried out. thus a high degree of concentration may be necessary before Effective mechanical disruption can help offset the need some types of extraction can begin. can be removed from their environment at total solids (2) physically contact the lipid material.. While many terrestrial feedstocks penetrate through the matrix enclosing the lipid material. extraction step by either converting whole algal biomass or Different methods can be used to disrupt the cell membrane by inducing the secretion of the desired product directly. While considerable is a promising development (Cravotto et al...5. which is dependant on the algal species and growth status. Laboratory-scale comparisons of process such as freezing. industrial-scale methods for extracting and separating of oils and lipids required for enabling biofuel production. little is known about the scale. and acid. to use elevated temperature and pressure processes that some algae-to-biofuels processes attempt to bypass the force the solvent into contact with desired biopolymers.. and microwaves and suggested that microwave disruption prior to solvent The basis for lipid extraction from algal biomass is extraction is the most efficient method (Lee et al. A shortfall of relevant information on efficient extraction bead mills (or bead-beating). Many effective of the biomass must be disrupted prior to extraction. and (3) solvate the >40%. 2010) and osmotic shock. prior to the application of the extraction solvents. To produce cells without disrupting cellular functions. autoclaving. To operations could affect extraction processes. sewage sludge with “focused pulse” sonication has been up separation challenges for extracted algal lipids. as well as the be successful. different Algal biofuel schemes that rely on the accumulation of harvest process operations (discussed in the Chapter 4) intra-cellular lipids are the focus of this discussion. and autoclaving of lipids and oils at larger-scale is limiting the algal-based (Mata et al. and Barberio. Assuming a system that requires extraction of oils and lipids from harvested biomass—as noted. 2008). though applications outside of analytical lipid extracts and preparation for conversion to biodiesel labs are unclear. 2010). that secrete products directly into the growth medium.

thereby improving downstream separations. it was found that when (2) the ability of a second less polar co-solvent to better applying direct transesterification using an acid catalyst match the polarity of the lipids being extracted. More. give relatively high hexane mixtures. 2007). In general. Fatty Acid Methyl Esters (FAMES) One example is the hexane/ethanol extraction co-solvent system (Grima et al. 1-butanol and ethanol) solvents have been tried proposed the direct transesterification of human milk and (Nagle & Lemke. and other lipid and non-lipid contaminants) nonpolar solvents weakening the association between the (Fajardo et al..g. solubilize.. It was found that the when selecting a co-solvent system to extract lipids are: entire reaction could be shortened to 10 minutes if the mixture was incubated at 100˚C under a sealed cap. The lipids mainly separate to the extraction (Lewis et al. but never more than 90% of the Bligh and recoveries of volatile medium chain triglycerides. lipids: hexane/isopropanol for tissue (Hara & Radin. chloroform. acetyl chloride) hydroalcoholic) that partition the extracted lipids into directly to the biomass sample and followed with heating the nonpolar hexane (Fajardo et al. making it more difficult for less polar solvents such microalgae (Nagle & Lemke.... 2000). The sequence of solvent addition can also affect two immiscible layers. despite the added added. biomass in a modified approach to include hexane in the reaction phase in order to avoid a final purification These results suggest that the two most important criteria step (Rodriguez-Ruiz et al. Consequently. water (which is not miscible with chloroform) is marine tissue that contained more than 2% lipids but added to the co-solvent mixture until a two-phase system worked well for samples that contained less than 2% develops in which water and chloroform separate into lipids. pure alcohol solutions of eliminate the need to use antioxidants to protect unsaturated greater carbon length (such as butanol) have not compared lipids. these findings suggest that membrane may be necessary. and hexane/isopropanol for lipids.e. has been promoted because hexane and alcohol will It is also noteworthy that the extraction efficiency was not readily separate into two separate phases when water is improved (when water was added first). This method was applied to dried microalgal well against the hexane/ethanol co-solvent system. at 100˚C for an hour under sealed cap would increase applications using pure alcohol (ethanol and 1-butanol) fatty acid concentrations measured (as compared to performed similarly. In these applications. the unsaponifiable lipids such as pigments. They explained lipids (i. hexane/ethanol for microalgae suggest that the water will form a solvent shell around the (Cartens et al. this approach achieved by adding both hexane and water in proportions suggested that a one-step reaction that added the alcohol that create a two phase system (hexane and an aqueous (e.Organic Co-solvent Mixtures Application of Organic Two-Solvent Systems for The concept of like dissolves like is the basis behind the Lipid Extraction from Microalgae: earliest and well-known co-solvent extraction procedure Iverson et al. agitation in the form of sonication or additional methanol. ethanol and Direct Transesterification of Lipids into isopropanol) have been suggested in place of methanol.. 1990). and extract the lipids.e. the efficiency of the reaction increased when a second “less polar” solvent such as To avoid the use of elevated temperature and pressure to diethyl ether or toluene was mixed with the methanol to push the solvent into contact with the analyte (at the cost modify the polarity of the reaction medium (Carvalho and of a very high input of energy).. 1990). 5. and Dyer co-solvent method. In general. dimethyl sulfoxide/petroleum ether for yeast extraction systems applied to “wet” biomass because they (Park et al. prior to their dissolution in the of co-solvents have been proposed for the extraction of monophasic system of water. less volatile and toxic alcohols (e. methanol) and acid catalyst (e. and methanol.. Similarly. In other cases. other combinations lipids and cell structure. single alcohol To increase analytical efficiency.e. These important results have a key impact on liquid phase 1978). it has been demonstrated that the extraction of lipids was significantly more efficient when solvents were added in order of increasing polarity (i. Starting from freeze dried chloroform layer and can then be recovered for analysis. 2007). Separation is then improved recovery of fatty acids. 2000).g. and then water) (Lewis. 1994). 2005). (i. Extraction of Products from Algae   43 . 1998).g.. In general. Lepage and Roy (1984) (e.. biomass. 2007). (1) the ability of a more polar co-solvent to disrupt the cell membrane and thus make it sufficiently porous and Continuing efforts along this path.g.. their results in terms of initial contact of the biomass with lipoproteins. The hexane system as chloroform to contact. the alcohol adipose tissue without prior extraction or purification for is first added as the extracting solvent. Chloroform will extract more than just the saphonifiable methanol. 1959). disruption of the cell Malcata. (2001) found that the Bligh and Dyer method (Bligh and Dyer. After the extraction reaction is grossly underestimated the lipid content in samples of complete. if not slightly better than alcohol/ Bligh and Dyer co-solvent system).. chloroform. 1996).. acetyl chloride).

10 min).55% of the fatty acids extracted by rate of mass transfer and degree of solvent access the Folch method in the first solvent extraction cycle.. By using solvents technique on 1 . the issue of solvent access to the material biomass. using the process. and optimize the energy cost. Hejazi et al. Extraction of Products from Algae   . being extracted is as important as the miscibility of the analyte in the solvent. Compressed gas is used to purge the different organic phases indicated that cells remained sample extract from the cell into a collection vessel. It has been proposed a technology that proposes to use solvents such as decane for the extraction of liquid extracts (Richter et al. (Denery et al. viable and active in the presence of organic solvents with a log P (octanol ) > 6 and that β-carotene can be extracted ASE is applicable to solid and semi-solid samples that more easily than chlorophyll by biocompatible solvents. The performance of ASE extraction was compared solvents (i. break down the cell membrane.the effectiveness of the second co-solvent (i. and showed that higher or equal extraction efficiencies remain largely bench-scale methods that are difficult (compared to traditional solvent technology) could be to scale up to industrial processes due to the actual achieved with the use of less solvent and shorter extraction solvent toxicity and the low carrying capacity of the times. 2006). and time have been optimized lipids coupled with its miscibility with methanol.30 g samples of dried biomass. and dodecane in the presence of live microalgal cells. Denery and coworkers optimized the extraction of carotenoids from Dunaliella salina All the preceding co-solvent systems.. to all pores within the biomass matrix and. Denery et al. ASE can also be applied to remove co.e. the polarity) to values that match the lipids. extracted 85 . What remains unclear is the effectiveness of such • the elevated pressures can reduce the dielectric an approach at large scale in terms of how to handle constant of an otherwise immiscible solvent (and large amounts of biomass. can be retained in the cell during the extraction phase (using a solvent front pumped through the sample at the This work has served as the basis for the development of appropriate temperature and pressure). ASE of higher hydrophobicity. This observation is a key driving force behind the consideration of solvent extraction Selective Extraction systems at elevated temperature and pressure. free fatty acids.e. 2004). concentrated for the extraction of triglycerides without 1998). 2009). 1996). If this process can be 44 5.. the effect of the solvent on uses organic solvents at high pressure and temperatures the membrane decreased and the extraction efficiency above their boiling point.. noteworthy in the context that ASE by definition uses non-aqueous solvents and therefore. In general. (2002) proposed the two-phase system of aqueous and organic phases for the selective extraction of carotenoids from the microalgae Dunaliella salina.. depending on the solvent. it is only efficient on biomass samples to that of traditional Folch method for microalgae grown containing less than 2% w/w lipids). a solid sample for both chlorophyll and β-carotene decreased as well... example. extraction temperature..200˚C) and pressure phase system of octanol and water. the cells can be returned extractable material from various processes. The latter is also polarity of the lipids (Herrero et al. is enclosed in a sample cartridge that is filled with an By applying a measurement of solvent hydrophobicity extraction fluid and used to statically extract the sample based on the partition coefficient of the solvent in a two- under elevated temperature (50 . must use dried Consequently. 1996. For to fractionate lipids from biological samples (ref). 2004) and lipids from microalgae (Schäfer. Their Accelerated Solvent Extraction observations were that solvents with lower hydrophobicity Accelerated solvent extraction (ASE) was first proposed reach critical concentrations more easily and in the in the mid 1990s (Richter et al. on dairy manure effluent (Mulbry et al. reaction ASE is more efficient if extracting solvent. however. In addition to improving yields and reducing loss of cell viability and extraction of membrane-bound extraction time.3000 psi) conditions for short time periods and activity tests of Dunaliella salina in the presence of (5 . sample-solvent medium) depends upon its ability to solubilize the target ratio. screening viability (500 . and production of triglycerides for biofuels production. separate out desirable by analogy. Accordingly. The single solvent systems at elevated temperature and ASE . a step that also requires energy input. to selectively to their original bioreactor for continued growth and extract polar compounds from lipid-rich samples.95% pressure have gained favor for two principle reasons: of the fatty acid content in the harvested microalgae • the elevated temperature and pressure increase the compared to 44 . Conceptually. some have proposed a modified technique to “milk” oils or neutral lipids from algae using biocompatible solvents and applied sonication.

2006). the fluid returns to its original gaseous state while al. There is also the added benefit of solvent extraction of essential oils from plants (Reverchon et access into the biomass matrix that occurs at the higher al. Metzger and cooled back down to room temperature.g. as well as pigments (such as chlorophyll).” was initially applied to whole biomass 2005). attractive aspects is the use of water as the solvent. at temperatures point can be adjusted as can the time of the extraction. low toxicity.. In addition. lower costs of the extracting agent.e.. non-photosynthetic algae are grown using will also require a significant cooling system to cool sugars as energy source and using standard industrial the product down to room temperature to avoid product fermentation equipment (Barclay et al.. products miscible Largeau. However. 2005). liquid feeds (Reverchon et al. Some degradation. 2006). The temperature and pressure above the critical liquefaction) is based on the use of water.9 bar). ease of scale-up.. Supercritical extraction is often employed in batch mode. one of the more to biodiesel (see Chapter 6 for more detail). as with accelerated solvent Heterotrophic Production extraction. 2006).. and chemical inertness (Luque de Castro et al.applied to microalgae slurries with suspended solid extraction techniques have been used in the commercial concentrations as low as 1 wt%.. The technique. is that water. the ability to 5. of the more important advantages described for subcritical economical production of biofuels from oleaginous water extraction include shorter extract times. Some pressures of 241 . Lipids have been selectively extracted at the high temperature and pressure become immiscible from macroalgae at temperatures of 40 . this method may provide extraction of substances from solid substrates. The basic premise to subcritical water extraction the extracted product remains as a liquid or solid. sulfur Subcritical Water Extraction hexafluoride.. water. thereby eliminating the need for the dewatering step. whether grown has the effect of converting lipids. it has been applied for the selective extraction the supercritical fluid. via transesterification. scaling up the process that is developed mainly for analytical usage. One of the more attractive points to supercritical hemicellulose as a pretreatment prior to its use as a fluid extraction is that after the extraction reaction has fermentation substrate (Mok et. by the same issues of energy-intensive processing and and environmental compatibility (Herrero et al. The potential benefits of this approach are the power of fluids above their critical point (Luque de use of standard fermentation systems. 73.379 bar (Chueng. as the water is from microalgae (Mendes et al. pressure are lowered to atmospheric conditions. however. creating additional energy use challenges. In this 2006). methanol. 1994. 1999). however. Supercritical fluid avoidance of expensive extraction scheme(s). Use of methanol as the solvating fluid With respect to microalgae. nitrous oxide. which can moderate critical temperature of 31. becomes less polar and organic compounds are more soluble than at room Supercritical fluid extraction has been applied for the temperature. al.. private companies have engineered algae that secrete oil into the fermentation media that can be recovered and later refined into a biofuel. phototrophically or heterotrophically. ethane. 2007). More recently. Large-scale design scenario. 1994).. The majority of applications have used the extraction of lipids that cannot be transesterified. a unique avenue for the selective extraction of lipids caffeine from coffee beans. as well as n-butane and pentane (Herrero et Subcritical water extraction (also known as hydrothermal al.. as well as functional ingredients and lipids temperatures as discussed above. easily separated downstream once the temperature and functional ingredients from microalgae (Herrero et al. triglycerides) that excludes (Brunner. water extraction.. Extraction of Products from Algae   45 . for more than two decades suitable for biofuels (e. this approach significantly Supercritical Fluid Extraction reduces the capital and operating cost for an extraction Supercritical fluid extraction utilizes the enhanced solvating process. It can be processed using solid and compared to photosynthetic systems. 2006). the solvent and product can be of essential oils from plant matter (Eikani et al. originally termed “pressurized hot but the process can also be operated continuously (Brunner. been completed and the extracted material dissolved into however. but other fluids used have included ethane. and saponins from oil-seeds (Güçlü-Üstündağ et case.1˚C and pressure of be difficult to separate from the desired lipids.. e. under these conditions. is the difficulty with designing a system at Other methods for extraction and fractionation include large scale and the high-energy load required to heat the the production of oils using heterotrophic algae. 2001) .50˚C and at lower temperatures and can be easily separated. In this system up to subcritical temperatures.. 2005).g. and pressure high enough to keep the liquid state (Soto and Luque de Castro. just below the critical temperature. higher productivity Castro et al. CO2 because of its preferred critical properties (i. 2007).. 1999). 1999). A major constraint. higher microalgae via supercritical processing is challenged quality of extracts. 1992).

Pressurized fluid extraction of carotenoids from Haematococcus pluvialis and Dunaliella salina and kavalactones from Piper methysticum. 73(8). to achieve sustainable commercial-scale systems. 46 5. 898-902. 175–181. 2007.. However. Extraction of Products from Algae   .. Meager. 80(2). Increasing the temperature helps to disrupt the solute- matrix interactions and to reduce the viscosity and surface tension of the water. 501(2). Fajardo. Separation of Desired Extracts from Solvent Stream 5. A. Presence of Water Associated with the Biomass such as chlorophyll and non. 2004. S.. K. reduce the pressure and temperature requirements. Increased of sugar-based feedstocks. As discussed in access to the material being extracted is as important Section 4. Golmohammad. Pressurized low polarity water extraction of saponins from cow cockle seed. W. G. intracellular benchmark can be considered: the extraction process water can prove to be a barrier between the solvent per day should consume no more than 10% of the total and the solute. 67(1-2).. produced per day. & Abril. The presence of water can cause problems at both ends at larger scales. F.. Comparison between extraction of lipids and fatty acids from microalgal biomass. 735–740. Eikani. Balsevich.3. The logistics challenges faced by the nascent lignocellulosic pressure also helps to force the solvent into matrices industry. versa. Journal of Food Engineering. et al. Mechanical disruption can large-scale heterotrophic “algal-refinery” operations.H. 21-33. Journal of Food Engineering.. 5049–5059..). When present in the bulk Energy Consumption and Water Recycle solution. as Btu. J. Journal of the American Oil Chemists' Society. Supercritical fluids: technology and application to food processing. Improved extraction of vegetable oils under high-intensity ultrasound and/or microwaves. thereby improving the References Barclay.. 2007. Eicosapentaenoic acid from the marine microalga Phaeodactylum tricornutum. & Malcata. Carvalho. This is a does not suggest that this is an insurmountable barrier. water can either promote the formation of emulsions in the presence of ruptured cells or participate For sustainable biofuels production. Lipid-Fett. J.maintain the integrity of the fermentation catalyst and use contact between the solvent and the solute.X.M. M. Ultrasonics Sonochemistry. Journal of Applied Phycology.. G. Subcritical water extraction of essential oils from coriander seeds (Coriandrum sativum L. 6(2). the following in side reactions. 2005. F. et al. 2007. Lipid extraction from the microalga Phaeodactylum tricornutum. 619–630. 123-129.transesterifiable lipids. et al. Grima. 1994. J. In this context.R.. 53(13). et al. the issue of solvent energy load.. 2005. M. 15(5). Chief amongst is the logistical challenge of that would normally not be contacted by solvents under securing a sustainable biomass feedstock to supply to feed atmospheric conditions. E. Güçlü-Üstündag. 2008. & Mazza.R.2 Challenges Extraction processes can yield undesirable components. 109(2). 120–126. A. Brunner. Very little information is available on this critical The extraction process is affected by the choice of step that is necessary before converting the algal upstream and downstream unit operations and vice biocrude into finished fuels and products. 80(2). Journal of the American Oil Chemists' Society. 1996. 1994. Journal of Food Engineering. significant downsides pressure facilitates enhancing the transport of the solvent to this approach include many of the same feedstock to the analytes that have been trapped in pores. Heterotrophic production of long chain omega-3 fatty acids utilizing algae and algae-like microorganisms. Food Chem. Preparation of fatty acid methyl esters for gas-chromatographic analysis of marine lipids: Insight studies. Cravotto. 1025–1031.. Denery. 955–959. et al. G. Analytica Chimica Acta. At the cellular level.P. a preliminary look at the energy balance as the miscibility of the analyte in the solvent. Agric. 71(9). & Rowshanzamir. principal motivation behind the application of extraction but it does serve to highlight the need for innovation techniques at elevated temperatures and pressures. Cartens. J.

Hara, A. & Radin, N.S., 1978. Lipid extraction of tissues with a low-toxicity solvent. Analytical Biochemistry, 90(1), 420–426.
Hejazi, M.A. et al., 2002. Selective extraction of carotenoids from the microalga Dunaliella salina
with retention of viability. Biotechnology and Bioengineering, 79(1), 29-36.
Herrero, M., Cifuentes, A. & Ibanez, E., 2006. Sub-and supercritical fluid extraction of functional ingredients from different
natural sources: Plants, food-by-products, algae and microalgae A review. Food chemistry, 98(1), 136–148.
Iverson, S.J., Lang, S.L. & Cooper, M.H., 2001. Comparison of the Bligh and Dyer and Folch methods for
total lipid determination in a broad range of marine tissue. Lipids, 36(11), 1283–1287.
Lee, J. et al., 2010. Comparison of several methods for effective lipid extraction
from microalgae. Bioresource Technology, 101(1), S75-S77.
Lepage, G. & Roy, C.C., 1984. Improved recovery of fatty acid through direct transesterification without
prior extraction or purification. The Journal of Lipid Research, 25(12), 1391-1396.
Lewis, T., Nichols, P.D. & McMeekin, T.A., 2000. Evaluation of extraction methods for recovery of fatty acids
from lipid-producing microheterotrophs. Journal of Microbiological Methods, 43(2), 107–116.
Luque de Castro, M.D., Jimenez-Carmona, M.M. & Fernandez-Perez, V., 1999. Towards more rational techniques
for the isolation of valuable essential oils from plants. Trends in Analytical Chemistry, 18(11), 708–716.
Mata, T.M., Martins, A.A. & Caetano, N.S., 2010. Microalgae for biodiesel production and other
applications: A review. Renewable and Sustainable Energy Reviews, 14(1), 217-232.
Mendes, R. et al., 1994. Supercritical carbon dioxide extraction of hydrocarbons from the
microalgaBotryococcus braunii. Journal of Applied Phycology, 6(3), 289-293.
Metzger, P. & Largeau, C., 2005. Botryococcus braunii: a rich source for hydrocarbons and related
ether lipids. Applied Microbiology and Biotechnology, 66(5), 486-496.
Mulbry, W. et al., 2009. Optimization of an Oil Extraction Process for Algae from the Treatment of
Manure Effluent. Journal of the American Oil Chemists' Society, 86(9), 909–915.
Nagle, N. & Lemke, P., 1990. Production of methyl ester fuel from microalgae. Applied Biochemistry and Biotechnology, 24(1), 355–361.
Park, P.K., Kim, E.Y. & Chu, K.H., 2007. Chemical disruption of yeast cells for the isolation of
carotenoid pigments. Separation and Purification Technology, 53(2), 148–152.
Reverchon, E., Iol & Marco, A.D., 2006. Supercritical fluid extraction and fractionation of
natural matter. The Journal of Supercritical Fluids, 38(2), 146-166.
Richter, B.E. et al., 1996. Accelerated solvent extraction: a technique for sample preparation. Anal. Chem, 68(6), 1033–1039.
Rittmann, B.E., Krajmalnik-Brown, R. & Halden, R.U., 2008. Pre-genomic, genomic and post-
genomic study of microbial communities involved in bioenergy.
Rodriıguez-Ruiz, J. et al., 1998. Rapid simultaneous lipid extraction and transesterification
for fatty acid analyses. Biotechnology techniques, 12(9), 689–691.
Schäfer, K., 1998. Accelerated solvent extraction of lipids for determining the fatty acid
composition of biological material. Analytica Chimica Acta, 358(1), 69–77.
Soto Ayala, R. & Luque de Castro, M.D., 2001. Continuous subcritical water extraction as a useful
tool for isolation of edible essential oils. Food chemistry, 75(1), 109–113.
Virot, M. et al., 2008. Microwave-integrated extraction of total fats and oils. Journal of Chromatography. A, 1196-1197, 57-64.

5. Extraction of Products from Algae   47

6. Algal Biofuel Conversion Technologies
Potentially viable fuels that can be produced from algae products, and this conservation law must also hold
range from gaseous compounds like hydrogen and methane, true for the algae biorefineries of the future if they
to alcohols and conventional liquid hydrocarbons, to are to achieve significant market penetration.
pyrolysis oil and coke. Attractive targets for this effort,
however, are the liquid transportation fuels of gasoline, A large number of potential pathways exist for the
diesel, and jet fuel. These fuel classes were selected as the conversion from algal biomass to fuels. These pathways can
best-value targets because 1) they are the primary products be classified into the following three general categories:
that are currently created from imported crude oil for the
bulk of the transportation sector, 2) they have the potential 1) Those that focus on the direct algal production
to be more compatible than other biomass-based fuels with of recoverable fuel molecules (e.g., ethanol,
the existing fuel-distribution infrastructure in the U.S., and hydrogen, methane, and alkanes) from
3) adequate specifications for these fuels already exist. algae without the need for extraction;

The primary objective of this chapter is to summarize a 2) Those that process whole algal biomass
number of potentially viable strategies for converting algal to yield fuel molecules; and
biomass into replacements for petroleum gasoline, diesel, 3) Those that process algal extracts (e.g., lipids,
and jet fuel. When a fuel meets all customer requirements, carbohydrates) to yield fuel molecules.
it is referred to as “fit for purpose.” While a successful These technologies are primarily based on similar
fuel-conversion strategy will address the full range of methods developed for the conversion of terrestrial
desired fit-for-purpose properties (e.g., distillation range, plant-based oils and products into biofuels, although the
ignition characteristics, energy density, etc.), these desired compositional complexities of the output streams from
fuel characteristics are driven primarily by customer algae must be dealt with before these can be applied
requirements and are discussed later in Chapter 8. This effectively. Pros and cons of these pathways within each
chapter focuses on fuel conversion strategies from a variety of these categories are discussed below, and a summary
of perspectives to establish the current state-of-the-art, of each fuel-conversion technology is given. Inputs,
as well as identify critical challenges and roadblocks. complexity, cost, and yields are provided (where known),
and key barriers and RD&D opportunities are listed.
Several emerged the Algal Roadmap Workshop in
relation to conversion of algal feedstocks to fuels. These
are noted here to help establish a reasonable framework 6.1 Direct Production of
for the most promising concepts for algal biofuels.
Biofuels from Algae
• First, the feedstock, conversion process, and
The direct production of biofuel through heterotrophic
final fuel specifications are highly interdependent
fermentation and growth from algal biomass has certain
and must be considered together if an optimal
advantages in terms of process cost because it can
process is to be identified. As a result, accurate
eliminate several process steps (e.g., oil extraction)
and detailed feedstock characterization (including
and their associated costs in the overall fuel production
both composition and variability) is essential,
process. Heterotrophic growth also allows for maintaining
since this is an upstream boundary condition for
highly controlled conditions, which first could be oriented
the entire downstream fuel-conversion process.
toward biomass production and then oil production. Such
• Second, life cycle analysis of energy and carbon will a system can generate extremely high biomass (hundreds
be a key tool in selecting the preferred fuel conversion of grams per liter) and a high percentage of that biomass
technologies from those discussed in this chapter. as lipid (well over 50%). The system is readily scaled up
• Third, the greatest challenge in algal fuel conversion is and there is an enormous potential to use various fixed
not likely to be how to convert lipids or carbohydrates carbon feedstocks (which would bring down the cost of
to fuels most efficiently, but rather how best to use production). These approaches are quite different from
the algal remnants after the lipids or other desirable the usual algal biofuel processes that use algae to produce
fuel precursors have been extracted. All of the biological oils which is subsequently extracted and used as
petroleum feedstock that enters a conventional a feedstock for liquid fuel production, typically biodiesel.
petroleum refinery must leave as marketable There are several biofuels that can be produced directly
from algae, including alcohols, alkanes, and hydrogen.

48 6. Algal Biofuel Conversion Technologies  

Alcohols In addition to ethanol, it is possible to use algae to produce
other alcohols, such as methanol and butanol, using a
Algae, such as Chlorella vulgaris and Chlamydomonas
similar process technology, although the recovery of
perigranulata, are capable of producing ethanol and other
heavier alcohols may prove problematic and will need
alcohols through heterotrophic fermentation of starch
further R&D. The larger alcohols have energy densities
(Hon-Nami, 2006; Hirayama et al., 1998). This can be
closer to that of gasoline but are not typically produced
accomplished through the production and storage of
at the yields that are necessary for commercial viability.
starch via photosynthesis within the algae, or by feeding
sugar to the algae directly, and subsequent anaerobic
fermentation of these carbon sources to produce ethanol Alkanes
under dark conditions. If these alcohols can be extracted In addition to alcohols, alkanes may be produced directly
directly from the algal culture media, the process by heterotrophic metabolic pathways using algae. Rather
may be drastically less capital- and energy-intensive than growing algae in ponds or enclosed in plastic
than competitive algal biofuel processes. The process tubes that utilize sunlight and photosynthesis, algae can
would essentially eliminate the need to separate the be grown inside closed reactors without sunlight. The
biomass from water and extract and process the oils. algae are fed sugars, the cheap availability of which
is a key consideration for cost-effective production of
This process typically consists of closed photobioreactors biofuels; these sugars are themselves available from
utilizing sea-water with metabolically enhanced renewable feedstocks such as lignocellulosic biomass,
cyanobacteria that produce ethanol or other alcohols while in a pressure and heat-controlled environment. This
being resistant to high temperature, high salinity, and process can use different strains of algae to produce
high ethanol levels—previous barriers to commercial- different types of alkanes; some algae produce a mix of
scale volumes (Hirano et al., 1997). There have been hydrocarbons similar to light crude petroleum. These
reports of preliminary engineered systems, consisting of alkanes can theoretically be secreted and recovered
tubular photobioreactors (Hirano et al., 1997). One key directly without the need for dewatering and extraction,
aspect of the system is that a source of cheap carbon, such but more often are associated with the algae and thus must
as a power plant, is typically used to supply CO2 to the be recovered through dewatering and extraction. With
bioreactors to accelerate the algae growth. An example further processing, a wide variety of fuels can be made.
of this process technology links sugar production to The process of growing the algae heterotrophically may
algal photosynthesis. There are claims that this process present some advantages over typical photoautotrophic-
may consume more than 90% of the system’s CO2 based technologies. First, keeping the algae “in the dark”
through photosynthesis, wherein a portion of the carbon causes them to produce more alkanes than they do in the
in these sugars is converted into ethanol. The ethanol presence of sunlight. While their photosynthetic processes
is secreted into the culture media and is collected in are suppressed, other metabolic processes that convert
the headspace of the reactor, purified, and stored. sugar into alkanes can become active. Second, some have
shown the growth rate of the algae to be much higher
This technology is estimated to yield 4,000 - 6,000 gallons than traditional methods (Xu et al., 2006). This is possible
per acre per year, with potential increases up to 10,000 because instead of getting energy for growth from sunlight,
gallons per acre per year within the next 3 to 4 years the algae get concentrated energy from the sugars fed into
with significant R&D. It is theoretically estimated that the process. These higher cell concentrations reduce the
one ton of CO2 is converted into approximately 60 - 70 amount of infrastructure needed to grow the algae, and
gallons of ethanol with this technology. With such yields, enable more efficient dewatering if is actually required.
the price of captured CO2 becomes significant, and may
require a price less than or equal to $10 per ton to remain Using algae to convert cellulosic materials, such as
cost-competitive. Further breakthroughs that enable more switchgrass or wood chips, to oil may have an advantage
efficient production systems and the development of new over many other microorganisms under development
process technologies may be critical in terms of long-term for advanced biofuel production. When lignocellulosic
commercial viability. Scaling of these systems to large- biomass is pretreated to allow for enzymatic hydrolysis
scale commercial biorefineries will also require significant for production of sugars, many toxic byproducts are
advances in process engineering and systems engineering. released including acetate, furans, and lignin monomers.
Metabolic pathway engineering within these algae, enabled In most other processes, these toxic compounds can
by metabolic flux analysis and modern genomics tools, may add process costs by requiring additional conditioning
further help in producing a commercially viable organism.

6. Algal Biofuel Conversion Technologies   49

McDermid & Stuercke. 2003. typical biomass throughout the country. improvement in hydrocracking. biohydrogen) substance by heating. algae: pyrolysis. biological hydrogen production can be considered a viable 2006).steps or the concentration of biomass hydrolysate There are four major categories of conversion in the conversion step. and supercritical processing (Exhibit 6. and the ground feedstock is quickly heated to 350 . too low for conversion to biodiesel.1). Algal Biofuel Conversion Technologies   . products. is a relatively new process (Bridgwater. higher efficiency can be achieved development of advanced photobioreactors. can result in a wide range of and dark-fermentation (see Chapter 2). The thermochemical treatment of indirect biophotolysis. 2003). especially amenable to processing a diverse range of algae. 2004). Hydrogen The production of hydrogen derived from algae has Pyrolysis received significant attention over several decades. Macroalgae has specifically received some attention as a gasification feedstock and initial work shows that while A significant roadblock in using pyrolysis for algae some key differences exist as compared to terrestrial conversion is moisture content. and detailed side-by-side comparison was found in 2003). fast There are several challenges that remain before heating rates. such as mannitol. and competitive drainage of fuel cannot yet be produced directly by pyrolysis of algae.2 Processing of Whole Algae units and has no fiber tissue to deal with. feedstocks must be ground into fine particles. and the added benefit of being fuels. laminarin. it appears fast pyrolysis to create bio-oil. with some hydrotreating and not only on research advances. requirement for Pyrolysis is being investigated for producing fuel from bicarbonate binding at photosystem II (PSII) for efficient biomass sources other than algae. a electrons by oxygen in algal hydrogen production. though from algae. gasification. anaerobic digestion. 2006. Pyrolysis is the chemical decomposition of a condensed Biological production of hydrogen (i. with hydrogen production by accumulation of a proton reaction times of the order of seconds to minutes. photo-fermentation.. and the United States. certain species are suitable for gasification (Ross et must be performed upstream for the process to work al. For flash pyrolysis. Several pilot plants for fast pyrolysis of biomass have been built in the In addition to the direct production of biofuels from past years in Germany. depending on the reaction parameters. Pyrolysis has one major advantage over other technology. degradable alternative liquid called bio-oil can be produced. The bio-oil has an advantage that it can enter directly The future of biological hydrogen production depends into the refinery stream and. Even with the increased methods benefit from reduced costs associated with interest in converting biomass into liquid transportation the extraction process. Also. This is one area where algae have a major advantage over other biomass sources because it exists fundamentally in small 6. There are several reports on the pyrolysis of algae in the scientific literature (Demirbas.e. and moderate temperatures (Huber et al.. in that it is extremely fast. or other biomass. at least some level of dewatering is still required. It does not involve reactions with technologies provide a wide range of approaches to oxygen or any other reagents but can frequently take generate hydrogen.. It appears that pyrolysis will concentrations approaching 20% are reported in some not be cost-competitive over the short-term unless an species (Chu et al. social acceptance. and significant dehydration crops. the algae. 2007). Miao and Wu. 2008). efficiently.e. place in their presence. to other biomass sources due to uncertainties in the Lipid content of a variety of macroalgal species is typically ability to dehydrate the feedstock. competitive inhibition of photosynthetic hydrogen production by CO2. but algae. It is unclear exactly how much more difficult and fucoidin. produce a suitable feedstock for generating efficiency through genetically engineered algae and/or the standard diesel fuel. the whole algae can be processed into fuels bio-oil from pyrolysis is not a commercial product at the instead of first extracting oils and post-processing. 2003). gradient. Brazil. although the scientific literature. where finely on economic considerations. Although synthetic diesel photosynthetic activity. no comprehensive less than 5% of total dry weight (McDermid & Stuercke. 2004). i. These include the restriction of photosynthetic conversion methods. 50 6.500˚C for development of a robust hydrogen infrastructure less than 2 seconds. These current time (Bridgwater. but also by the so-called “flash pyrolysis” technology.. Polysaccharides. Liquid product yield tends to favor short residence times. represent the main macroalgae biochemical it would be to convert algae into a bio-oil compared feedstocks for conversion to liquid fuels (McHugh. including direct biophotolysis.. Algae may prove to be more technologies that are capable of processing whole resistant to these compounds and sugar conversion.

Unfortunately. FTS is also a relatively demonstrated that the bio-oil produced by pyrolysis mature technology where the syngas components (CO. 2007. 2004). CO2. since pyrolysis is already a Gasification of the algal biomass may provide an extremely relatively mature process technology. address the issue of availability easily transported throughout the upgrading process.inexpensive dewatering or extraction process is also Gasification developed. Yung et al.. 2006).. algal bio-oils from different feedstocks need to be carefully Additionally. there are also significant gaps in the information available about the specifications for Conversion of bio-syngas has several advantages over other converting algal bio-oil and the resulting products. 2009). Exhibit 6.. Another advantage is the Additionally. making detailed molecular composition of the resulting bio-oils. Srinivas et al. The methods. Algal Biofuel Conversion Technologies   51 . alcohol synthesis of the resulting syngas. Another research paper comparatively straightforward. the process more flexible. so the bio-oil may be more investment required.1 Schematic of the potential conversion routes for whole algae into biofuels Algae Anaerobic Supercritical Pyrolysis Gasification Digestion Fluids SynGas Liquid or Fischer Higher Alcohol Vapor Fuel Tropsch Synthesis Catalytic Catalytic Upgrading Upgrading Transportation Liquid MeOH Biogas Fuels . liquid fuels through a water-gas shift and CO hydrogenation (Okabe et al. and it of compounds and compositions depending on the is reasonable to expect that once water content is adjusted type of algae and upstream processing conditions for. it may have a different range relatively mature (Phillips. the gasification of algae to these biofuels would be (Zhang et al. bio-syngas is a versatile feedstock and it studied. Work also needs to be performed to understand the can be used to produce a number of products.Liquid or Hydrogen Gas Hydrogen Fuels EtOH. Additionally. Another area of interest is the thermochemical infrastructure.. H2. The synthesis While algal bio-oil may be similar to bio-oil from of mixed alcohols using gasification of lignocellulose is other biomass sources. and impurities) are cleaned and upgraded to usable algal growth conditions (Miao and Wu. 6. Balat. 2007. research is needed on the catalytic conversion possibility to integrate an algal feedstock into an existing of the resulting algal bio-oils. primarily that only incremental improvements will occur and a through Fischer-Tropsch Synthesis (FTS) or mixed breakthrough in conversion efficiency appears unlikely. it is expected flexible way to produce different liquid fuels. it is possible to create a wide optimal residence time and temperature to produce different variety of fuels with acceptable and known properties. etc. 2009. First and foremost. of algae can be tailored by carefully controlling the H2O. It may be possible to feed development of stabilizers for the viscosity of the bio-oil algae into a coal gasification plant to reduce the capital and acid neutralizing agents. 1994).

The tars cause coking of the shortened time scales. usually requires pretreatment.. algae may have a unique advantage balance. These technologies harness the high synthesis catalyst and any other catalysts used in the activity of water in subcritical environments that is capable syngas cleanup process and must be removed. First. A maximum oil yield of 64% using Botryococcus in this process. The first reactor was a low. it is necessary to determine the optimum conditions 52 6. of the feedstock on a • Fluidized-bed gasification and solvent tar removal dry ash free basis and has energy content that is comparable • Fluidized-bed gasification and to diesel and can be upgraded further. comparable to those of fuel oil. The tar-rich gas was then passed of algae to date.. it would be useful to leverage dewatering process with a regenerative heat exchanger. more research in this area is needed through an entrained-flow gasifier where it was reacted before it can become a commercially viable option.4 wt% produced an oil Research Centre of Finland (VTT). Even though FTS is a mature technology. with the exception of any upstream process steps that may be Direct hydrothermal liquefaction in subcritical water a source of contaminants which will need to be removed (defined as water held in a liquid state above 100oC by prior to reaching the FT catalyst. 2005). the most significant problem al. Another approach for tar-free syngas was liquefaction technique was more effective for extraction demonstrated in a pilot plant in Freiberg.330 mPas and a calorific value of Tar formation can be minimized or avoided via entrained. While this technology requires sub-millimeter a net energy producer from the overall process energy sized particles. • Entrained-flow gasification at high temperature defined as the ratio of heating values of bio-crude products and feedstock plus external heat input. Also. Neste Oil. Liquefaction of algae is two successive reactors. In the same flow gasification at high temperatures (Hallgren et al. The oil obtained at a reaction installed in a plant in Moissannes. There are reports subsequent thermal tar cracker (Goudriaan et al.. 2008). 2005). The a small size with other biomass sources and doing so algal biomass was processed by liquefaction at 300˚C.for dedicated biomass plants. This technology is a representation of the natural with FTS is the cost of clean-up and tar reforming. 2006). considered a promising technological approach but due temperature gasifier that broke down the biomass into to limited information in hydrothermal liquefaction volatiles and solid char. FTS tends to require applying pressure) is a technology that can be employed to production at a very large scale to make the process convert wet algal biomass to a range of liquid fuels (Patil et efficient overall. (Raffelt et al. Prior work in direct liquefaction of biomass was very A demonstration plant for gasification of wood chips active. Liquefaction of Dunaliella tertiolecta built in Finland in a joint venture of the Technical with a moisture content of 78. as high as 75%. and improve the process for indirect gasification of algae. since determine the feasibility of using the oxygen generated FTS is an exothermic process. with oxygen at high temperature. The key roadblocks to using FTS for algae are thought Liquefaction to be similar to those for coal (Yang et al. and there are a few reports that used algal biomass with catalytic cracking of the tar is currently being as a feedstock. 1995). Algal Biofuel Conversion Technologies   . there are still several areas that should be investigated and require R&D. report it was concluded that liquefaction technique was 1993).. Additionally. Germany. it is difficult to reach such braunii as a feedstock has also been reported... The main product of this liquefaction process is a “bio-crude” • Fluidized-bed gasification and catalytic reforming that typically accounts for 45% wt. it should be possible to use by algae for use in the gasifier to reduce or eliminate the some of the heat for drying the algae during a harvesting/ need for a tar reformer. Tars geological processes known to be involved in the formation have high molecular weight and can develop during of petroleum-based fossil fuels realized over greatly the gasification process. and yield of about 37% (organic basis) at 300˚C and 10 MPa Stora Enso. but certain species of catalyzed by sodium carbonate (Sawayama et al. It would be desirable to efficiency through economy of scale. The four of decomposing the algal biomass into smaller molecules basic mechanisms to deal with tar-related problems are: of higher energy density or more valuable chemicals. temperature of 340˚C and holding time of 60 min had a viscosity of 150 .. algae may not require pretreatment due to their inherent There have also been comparative studies that report the small size. ongoing syngas-to-ethanol research using cellulosic. Typically. 1995). of microalgal biodiesel than using the supercritical carbon built by Choren Industries GmbH. The pilot plant used dioxide (Aresta et al.. However. 36 kJ g−1. A solvent tar removal demonstration was (Minowa et al. 2000) that claim the thermal efficiency. France in 2006.

have been types of inputs to the conversion technologies must be shown to be very favorable for large-scale deployment. In for example. such as Laminaria hyperbore and simultaneous extraction and transesterification of vegetable Laminaria saccharina. and this be mitigated through the addition of a transesterification technique has been demonstrated to be extremely powerful conversion process.. It also remains to be seen whether the processing of whole algae in this The conversion of extracts derived from algal sources is fashion is superior. at less than 50˚C. Additionally. gal vs. further work should be focused on the using strains not optimized for lipid production.. $0.. the typical mode of biofuel production from algae. Several studies have been yield similar to that of the anhydrous process in the carried out that demonstrate the potential of this approach. The most common type of algal extracts under One economic analysis has been conducted based on a consideration are lipid-based. supercritical methanol in the United States (Hanssen et al. is to demonstrate are selective. which supercritical process to produce biodiesel from vegetable 6. with a methane concentration of 65% (Vergara- al. 2007). a consequence of temperature and pressure conditions where algae are grown under uncontrolled conditions used in this study. Similar results have been Fernández et al. however. Algal Biofuel Conversion Technologies   53 . it must be demonstrated that this process can product stability and quality. it has been demonstrated that this extraction. If this approach can be modified observed for supercritical methanol processing of vegetable for the use of microalgae. 2008. Supercritical fluids The clear immediate priority.3 Conversion of Algal Extracts be needed to avoid saponification. conversion of vegetable oils. therefore. Additionally. In this process variant. supercritical tolerate the complex compositions that are found with fluids can be used on whole algae without dewatering. in place. $0.. 2008). there are no organic solvent applied in the processing of algae. and fuel conversion. in particular. it is envisioned as being applicable for the processing biofuel production. an obvious and critical link between the type of extraction process used and the product composition. transesterification. 1987). e.4 ml/g-d of biogas can be realized using a two- efficiency. Vieitez et al. including drying. the decomposition of fatty acids is the main stage anaerobic digestion process with different strains factor that limited the attainable ester content (Vieitez et of algae. either whole or its residues in the extract or spent biomass (Demirbas. fundamental and exhaustive understanding of the different at least in the case of vegetable oil processing. is an interesting mode of gaseous oils. and efficiency. The production of biogas from the anaerobic digestion 2008). and one that receives scant attention of algae. lower reaction temperatures... 2009). While the occurrence of A recent study indicated that biogas production levels water in the reaction medium appears as a factor in process of 180.. and as such may be a process is capable of tolerating water. In the case of combined extraction and transesterification of algae. triacylglycerides. This supercritical transesterification approach can also be applied for algal oil extracts. and as such. it may be very effective oils (Hawash et al. It is. level that can be scaled to commercial production. cost.e. key obstacles that are responsible for the current high 2004). Although it has been only demonstrated for the of macroalgae. The supercritical extraction process can be coupled with a transesterification reaction scheme to enable a “one Anaerobic Digestion of Whole Algae pot” approach to biofuel production (Anitescu et al. unprocessed algae and that there is no negative thereby increasing the efficiency of the process..26/ and converting oils into biofuels (Demirbas.51/gal).. the overall algal biorefinery in the extraction of other components within algae (Mendes. Because decomposition was for situations like integrated wastewater treatment. in terms of yield.Supercritical Processing oils in one step using alcohols (Anitescu et al.g. It was found that the processing cost of the proposed Supercritical processing is a recent addition to the portfolio supercritical technology could be near half of that of the of techniques capable of simultaneously extracting actual conventional transesterification methods (i. 2008). 2009). There is to the transesterification of the algal oil extracts. impact due to the presence of other small metabolites. further study will also 6. thus providing high purity and product that these supercritical process technologies can be concentrations. with a conversion cost-effective methodology. theoretically possible Supercritical fluid extraction of algal oil is far more efficient that if the other upstream algal processing costs could than traditional solvent separation methods. The use of or ethanol is employed as both the oil extraction medium this conversion technology eliminates several of the and the catalyst for transesterification (Warabi et al. ensuring maximum particular. 2006). a The economics of supercritical transesterification process. raw. In the case of catalyst-free supercritical ethanol costs associated with algal biofuels. could become cost-competitive with fossil fuels. 2009). with similar yields and efficiencies at a Extraction is efficient at modest operating temperatures. oil extract. effect of milder process conditions.

This section discusses water and free acids. therefore. mitigate saponification chemical. Olefin Diesel Diesel Aromatics 54 6. the reaction soluble bases by liquid acid catalysts such as H2SO4. there are other solution and mixing well. and therefore. The apparent higher activity of certain standard” in the conversion of vegetable oils into biodiesel HPAs with respect to polyoxometallates of higher (Hossain et al. 2006).. strength resulted in lower pretreatment temperatures. Algal Biofuel Conversion Technologies   . catalyzed methanol approach.2). which is simply a process of such as H3PW12O40/Nb2O5. have been shown to lower the displacement of an alcohol group from an ester by required temperatures and decrease the reaction times another alcohol (Demirbas. Cao et al. 2008). (Ataya et al. and catalytic processes that can and emulsification. 2007).can be converted into biodiesel. it has been shown that One recommended research focus would be to further transesterification of algal oil can be achieved with ethanol develop these homogeneous catalysts to tolerate the and sodium ethanolate serving as the catalyst (Zhou & contaminants expected to be present in algal extracts. In addition to the classic base. Recently. enhancing the product recovery be employed to convert algal extracts. it was can be performed via catalytic or non-catalytic reaction shown that HPA-catalyzed transesterification of vegetable systems using different heating systems that are required oil achieves higher reaction rates than conventional to initiate the reaction. 2008).. HCl is accelerated and requires shorter reaction times. Biodiesel is then separated processing variants that appear promising. 2008). Kerosene. they are not currently preferred due to their lower activity than the conventional transesterification Chemical Transesterification alkaline catalysts. This technology is relatively mineral acids due to their higher acid strength (L. As a or H3PO4 is also considered an attractive alternative as result. Transesterification (Alsalme et al.. In triacylglycerols extracted from algae to FAMEs (fatty order to compensate for this. acid methyl esters). Boocock. 2008). heteropolyacids (HPA). 2008). An alternative from the ether by a vaporizer under a high vacuum.. (Exhibit 6. heating system that can be used to enhance the kinetics of transesterification involves the use of microwaves (Refaat Another route is found in acid-catalyzed transesterification & El Sheltawy. biochemical. generally required as a result. 2008.. 2009).2 Schematic of the various conversion strategies of algal extracts into biofuels Algal Oil Extracts Chemical Enzymatic Catalytic Transesterfication Conversion Cracking Liquid or Biodiesel Vapor Fuel (ME) Catalytic Catalytic Upgrading Upgrading Gasoline. The products of these reactions are typically separated by adding ether and salt water to the In addition to alternative catalysts.. Xu mature and has been demonstrated to be the “gold et al. Though acid catalysts have these advantages. The replacement of is carried out in the presence of microwaves. Diesel. a drastic reduction in the quantity of co-products the acidic catalysts are less sensitive to the presence of and a short separation time are obtained (Lertsathapornsuk Exhibit 6. Higher temperatures and longer reaction The transesterification reaction is employed to convert times are. When the transesterification reaction reactions (Wahlen et al.

Although enzymatic approaches have become converting lipids into a suitable diesel surrogate. This cavitation provides simultaneously reuse. catalysts may be used to enhance the enzymatic biodiesel production has several advantages: impact of microwave irradiation (Yuan et al. 2008). converting carbohydrates into conventional processes (Svensson and Adlercreutz. novel enzymes with desired characteristics that are more suitable for industrial applications (Guncheva et al. to their corresponding esters but have drawbacks such as being energy-intensive.. Although very effective and relatively economical.. but (Lopez-Hernandez et al. much R&D is needed in the discovery. process must be able to function in the presence of these However. in many cases. One potential solution to this is the enable efficient biocatalytic processing. These preliminary results indicate that that a co-solvent mixture may be critical in defining microwave processing may be cost-competitive with the the optimal reaction medium for the lipases. It is capable of producing these reactions in a variety of estimated that industrial-scale ultrasonic devices can allow environments and on different types of oil feedstocks for the processing of several thousand barrels per al. and energy input (Kalva et al.. it has been noted 6. these catalysts still require purification One critical area that needs to be addressed is the solvent and removal from the product stream. have been proven as a viable means of triacylglycerols for biodiesel production (Su and Wei. one of the most transesterification process (Armenta et al. Enzyme immobilization may also play Biochemical (Enzymatic) Conversion a key role in developing an economic method of Chemical processes give high conversion of triacylglycerols biocatalytic transesterification (Yamane et al. Use of biocatalysts cell walls. There have been of the two types of sites in the overall reaction in terms some recent reports of using a solvent engineering of efficiency and cost.. Lewis acid catalysts.. optimizing specific enzyme activity to (lipases) in transesterification of triacylglycerols for function using heterogeneous feedstocks. production. Other important issues that need further exploration and require removal of alkaline catalyst from the product are developing enzymes that can lyse the algal and treatment of alkaline wastewater. Acid and basic catalysts and the enzymes used in the downstream conversion could be classified as Brönsted or Lewis catalysts. both types of sites could be solvents to varying degrees to enable cost-effective present and it is not easy to evaluate the relative importance biofuel production (Fang et al. 2008). the reaction mixture to produce and collapse bubbles (c) no catalyst regeneration steps are needed for lipase constantly. 2008). This work more mature conversion processes currently available. Thus significant roadblocks to demonstrating the validity of using an ultrasonic reactor for biodiesel production this approach lies in the conversion of algal oil extracts drastically reduces the reaction time. and optimization of enzymes that are catalyzed transesterification (Stavarache et al.. Algal Biofuel Conversion Technologies   55 . ultrasonic waves cause (b) high reaction rates and conversion can be obtained. 2008). 2007). difficulty in removing the glycerol. 2008). and 2008). time-consuming batch process used in traditional base.. sugars. The transesterification catalysts presented above are These factors must be addressed before a commercially very strong and relatively mature in the field of biofuel viable biochemical conversion process can be realized.. Again. they have not been demonstrated at large scale mainly due to the relatively high price of lipase and its short operational life caused by the negative Catalytic Cracking effects of excessive methanol and co-product glycerol. at a commercial scale and at competitive prices. In the ultrasonic reactor method.. 2005). increasingly attractive. In particular. defining biodiesel production addresses these problems and necessary enzyme reactions (cell wall deconstruction offers an environmentally more attractive option to the and autolysin). which increases and temperature tolerance of the enzymes in order to the overall costs. 1998). 2003). 2008. inexpensive (McNeff et al. The presence of development of immobilized heterogeneous and/ solvents is sometimes necessary to enhance the solubility or homogeneous catalysts that are very efficient and of the triacylglycerols during the extraction process. 2006). Hence the process of transesterification can run inline rather than using the To that end. as with other approaches. reaction temperatures. and (d) the operational stability of the catalyst is the mixing and heating required to carry out the high. catalyzing nucleic acid hydrolysis. Liao et al. Bioprospecting for will require further innovation to reach production levels the enzymes in extreme environments may produce sufficient for massive and scalable biofuel production. (a) both the negative effects caused by excessive methanol and co-product glycerol can be eliminated completely. 2007). such as method to enhance the lipase-catalyzed methanolysis of AlCl3 or ZnCl2. indicates that the use of this co-solvent mixture in the In addition. engineering. converting triacylglycerols into fatty acid methyl esters..

and diesel are generally described as amounts of free fatty acids and water. This catalyst is also more active than HTiNbO3. using autoignition characteristics (as measured by octane number HTiNbO3 as the catalyst. begun to explore the conversion of vegetable oils and waste animal fats into renewable fuels. It is worth noting that the Double-metal cyanide Fe-Zn proved to be promising petroleum industry began by developing a replacement catalysts resulting in active transesterification of oil. Many of the catalysts cracking.. such as tehtrahydrofuran. reducing the concentration the same performance specifications as the petroleum- of free fatty acids in non-refined oil or in used oil. and Jet Fuel efficiencies of up to 98% (Soriano et al. obtained in a fixed-bed reactor at 200˚C and 35 bar. oil industry has begun to cast a wider net and has spent 2006). As noted in chapter 8. and x is an and process steps. A major characteristic of petroleum-derived catalyst examples include MgO. be completed before these catalysts are fully understood. CaO. and insoluble. straight chain alkanes are poor starting materials for gasoline because they provide low octane numbers. M a niobium atom or a tantalum atom. active catalysts during transesterification. There remain significant can be used for the production of typical fuels without fundamental studies and unanswered questions that must disruptive changes in processes or infrastructure. which translates to very costly plant design of those operations already in place: thermal or catalytic and construction requirements. the with TiVO4 being the most active (Cozzolino et al. but have essentially in the esterification reaction. and cloud point. From a refinery’s At these temperatures. All of the processes that take place in a modern petroleum refinery can be divided into two categories. The fuel products are a which A represents a hydrogen atom or an alkaline metal mixture of components that vary based on input stream atom. Fatty acids are well suited to conversion to diesel and jet fuel with few processing steps. and now it is apparent that it is beginning to These catalysts are Lewis acids. and Al2O3. initial and final boiling point. One particular concern is the stability and longevity of Various refiners and catalyst developers have already the catalysts in a representative reaction environment. they can be used even with oils containing significant Gasoline. The catalysts are active feedstock. the process development but must resist poisoning and the feedstock is considered fungible with petroleum and leaching of active components. jet fuel. 56 6. Algal Biofuel Conversion Technologies   .60 bar). in comparison to the homogenous catalyst at lower therefore. On the other hand. the process pressure is high perspective. a great deal of resources developing additional inputs producing the same yields with lower residence times. temperatures of about 170˚C). fuels is high energy content which is a function of a near-zero oxygen content. separation In another example. hydrophobic (at reaction return to biological feedstocks to keep the pipelines full. presented above seem to be good candidates for industrial and catalytic structural isomerization. probably due to the “renewable” or “green” if it is derived from a biological hydrophobicity of their surface. In this way. 2009). catalysts derived from the titanium and modification of the components in crude oil to yield compound possessing the general formula ATixMO. demanding additional isomerization steps or high octane blendstocks. This process has already provided the renewable jet fuel blends (derived from oils obtained from jatropha and algae) used in recent commercial jet test flights. the predominant feedstock for the industry is crude oil. Other based analog. A typical diesel must meet a multitude of performance specifications FAME yield of 91% and glycerol yield of 91% were that include volatility. and they are better defined by their integer not greater than 7. Typical biological molecules One of the most difficult challenges is finding an ideal have very high oxygen contents as compared to crude oil. The catalysts obtained are stable and molecules.. Conversion to Renewable Diesel. such as biomass or plant oil. is largely a process of eliminating oxygen and temperatures than the ones currently used (~220 . Moreover.240°C). Vanadate metal compounds or cetane number). flash point. Although are stable. maximizing the final energy content.The presence of a co-solvent. and give high glycerol yield with high activities. jet fuel. such as oil shale and tar sands. catalytic hydrocracking and hydrotreating. can play a vital role in achieving high conversion Gasoline. were employed in vegetable oil performance specifications than by the sum of specific transesterification. the ideal conversion process would make use (40 . for whale oil. in an assortment of end products. heterogeneous catalyst that has comparable activity Conversion of biological feedstocks to renewable fuels. gasoline.

. 6. This includes the anaerobic digestion of algal remnants The primary barrier to utilizing algae oils to make to produce biogas. nitrogen from extracted proteins. Catalysts in any recoverable polysaccharides into biofuels. 1979).Algal lipids can be processed by hydrothermal treatment 6. Algal Biofuel Conversion Technologies   57 . carbon dioxide. current use have been optimized for existing petroleum feedstocks and have the appropriate specificity and activity Anaerobic digestion can be effectively used as a means to carry out the expected reactions in a cost-effective of producing biogas from algae and algal remnants after manner. and reduce double bonds to yield technology that derives benefit from every potential hydrocarbons. In particular.g. as well as the amount of are amenable to anaerobic digestion. contaminants to arrive at the most cost-effective process. Glycerin can be converted to propane input is the conversion of algal remnants after which can be used for liquefied petroleum gas. once H2 used. It will be desirable to tune catalysts such that the extraction (Ashare and Wilson. Refinery catalysts have also been developed to the algae has been harvested. little if any pretreatment function within a certain range of chemical components is required. conversion of algal feedstock into fuel. Referred to as hydrotreating.4 Processing of Algal (basically. this process will convert the carboxylic Remnants after Extraction acid moiety to a mixture of water. contains soluble nitrogen from the original algal proteins. There will also likely be a high metals (especially magnesium) from chlorophyll. It will amount of polysaccharides and other oligosaccharides be necessary to optimize both the level of purification of present in the algal remnants that are well suited for algal lipid as well as the tolerance of the catalyst for the traditional fermentation into ethanol and other biofuels. The liquid effluent and nitrogen heteroatoms) without becoming poisoned. a chemical reductive process). the attack on the oxygen-bearing carbon atoms will minimize organic fractions of the algae remaining after oil extraction the amount of CO and CO2 lost. and sulfur methane and 40% CO2 by volume. as well as the fermentation of renewable fuels is catalyst development. Crude algal oil may contain high levels of phosphorous the nitrogen can be recovered in the form of ammonia from phospholipids. and for recycle to the culture. The biogas product typically contains 60% found within the petroleum stream (e. or One other critical aspect in developing a conversion carbon monoxide. metals. In addition.

P. M. 1391-1399. Y. In Gaigneaux E.. Acidolysis of tripalmitin with oleic acid catalyzed by a newly isolated thermostable lipase. Demirbas.M. F.. support and type of precursor on the catalytic performances in the ODH of methanol to formaldehyde. Part A: Recovery. (2006).V. 1679-1693. and Bjerle. Di Serio.. R. J. Martens. Van der Wal. J. 1045-1052. (pp. 2006.. P. Symposium.M. 21–50. A... September 10-14. A.. Louvain-la-Neuve. M.E. 86(14-15). A. (1979)... Kenawi. 697-704. Hirayama.. (2008).. 349(1-2)... 338-49. (2008). and Ascophyllum nodosum and application of end products. Fuel Processing Technology. 1312-1325). Part A: Recovery. Biodiesel fuel from Jatropha oil via non-catalytic supercritical methanol transesterification. E.. (2005). Demirbas.R. and Tavlarides. and Ruiz. Energy Sources.A. Wang. Kozhevnikova..A. Hirano. 58 6.M. Bridgewater Anthony.].. Current Microbiology. 22(2-3). MA (USA). Tyrol.. Aresta. N. and Barrow. Education. Andersson. M. and El Diwani. Radchenkova. 3rd (1994).. Norazmi Mohamed.. S. 28(10).. Indergaard. and Xu. Energy Fuels. Kamal. Applied Catalysis A: General. G.. Biomass fast pyrolysis.. Tesser. M. Oxford. (Eds. 101(1). (2004). Anaerobic digestion of Laminaria spp.. (1993). E.. Dynatech R/D Co. Oestgaard. CO2 fixation and ethanol production with microalgal photosynthesis and intracellular anaerobic fermentation. and Ternan. V. A. A. Naber. (2009).L. A.. Hanssen.. O.References Alsalme. Cao. T.. E. Eloy.. 53(6). R. 933-940. Fuel. In (Ed) Bridgwater. Dube. E. (2006). COO-2991-42. Y. Hawash. Variables affecting the induction period during acid-catalyzed transesterification of canola oil to FAME. Lv. (2009). Bos. Ataya. 923-927. G. Fatty Acid Composition of Some Malaysian Seaweeds. 579-582. UK: Blackwell Science. Energy Sources.V. and Barberio. B.. Analysis of digester design concepts. J. Adv. 9th Int. & Phang... Goudriaan. O. 88(3). Zaher. M..) Studies in Surface Science and Catalysis. Conf.. Demirbas. K.. and Environmental Effects.. I. X... 14(1). I. Int. Cozzolino. 1-13. J. L. Armenta. Pedersen. 17(1-2). Vinatoru. Dibenedetto. S. and Jensen. Fang. Biodiesel production from high acid value waste frying oil catalyzed by superacid heteropolyacid... [Ed. Biomass. J. Energy and Fuels.. A.. Proceedings of the Progress in Thermochemical Biomass Conversion. Thermal efficiency of the HTU® process for biomass liquefaction. (2000). Austria. Heteropoly acids as catalysts for liquid- phase esterification and transesterification. (2006). 21-27. and Kambourova. Gaigneaux. S. L. (1997). Thermochem.. Biotechnology and Bioengineering. and Zhu. R. Devillers.J. M.. Zhiryakova.(2006). De Vos. A newly isolated organic solvent tolerant staphylococcus saprophyticus M36 produced organic solvent-stable lipase. Anitescu.. A.. 17-22. 31(2).. X. and Santacesaria.. S. 22(2). W. and Environmental Effects... A. 83-103.. 510-515.. Biomass Convers. W. A. F. Scientific Bases for the Preparation of Heterogeneous Catalysts Proc. M. A. (2008). W. 84(11). Energy-Oxford. 170-176. Baevre.. F. Chen. & Wilson. and Kozhevnikov. Kralovec. Burja. Zhai. C. 93-100. Utilization. and Zeevalkink. Biodiesel from waste cooking oil via base-catalytic and supercritical methanol transesterification.. (1987). S. Wang. 50(4). Chu.. Utilization of macro-algae for enhanced CO2 fixation and biofuels production: Development of a computing software for an LCA study. 163-168. 2000.E. N. Z. High temperature gasification of biomass in an atmospheric entrained flow reactor. Malaysian Journal of Science. Integrated technology for supercritical biodiesel production and power cogeneration. Ashare. Belgium. Ding. Wang. G. Hallgren. Hermans. 162. F. F. S. Lu.. Pap.. Energy Conversion and Management.A. Ueda.. 22(1). A. (2009). D. Guncheva. Production of biodiesel from algae oils. M. Deshpande. Oily products from mosses and algae via pyrolysis. Science and Technology...E. Bie. Utilization. M. Supported vanadium oxide nanoparticles: Effect of preparation method. J.. Journal of the American Oil Chemists’ Society. F.E.. F. M. Z.. M. 8(2). Y.. 679-685. 22(2). F. (2007). P. Liu. Meeting Date 1993. A.A. Jacobs. H. 129-132.. (2008). 137-142. Sep. G. Li. Thermal Science. S. Boerefijn. 85(2). Sustainable transportation fuels from biomass materials.M. J. D. Cambridge. and Ogushi. A. E. Transesterification of fish oil to produce fatty acid ethyl esters using ultrasonic energy. Journal of the American Oil Chemists’ Society. (2003). Van de Beld. Rev.F. (2008). Algal Biofuel Conversion Technologies   . Energy. Balat.

. S. Y.. and Giselroed. F. 110(1). S.. P. (2003). R. Food and Agriculture Organization. 189-213).. 85-93. T. Industrial and Engineering Chemistry Research. Stavarache. Lipase-catalyzed transesterification of medium-chain triacylglycerols and a fully hydrogenated soybean oil. M. 131(1-3). Ultrasonics Sonochemistry.G. 25(21). 808-828. (2006). A. Krohn. Applied Biochemistry and Biotechnology. Sawayama. and Moholkar.. A. S. 4(3). Steinhardt... 88(3). and Krisnangkura. Lertsathapornsuk.. McNeff. C365-C372. Inc. A. A. 171-176. and Liu. Fuel.. Chlamydomonas perigranulata. V. Gyberg. The BTL2 process of biomass utilization entrained-flow gasification of pyrolyzed biomass slurries. (2006). M. Soriano. and Sanjay M. Ogushi.. 4(4). Nowlan... Patil. & Stuercke.. J.W. Srinivas. A. Journal of Biotechnology. V. A. L. Biodiesel synthesis via homogeneous Lewis acid-catalyzed transesterification. Dote. Biodiesel fuel production from algae as renewable energy. Raffelt.). B. (2007). K. H.Hirayama. 11(4). C. WSEAS Transactions on Environment and Development. 657-660. Venditti. (1998). A... Jr. (2009). Y. 74. 153-164. Huber. Ueda. (Ed.Y...L. Application of solvent engineering to optimize lipase-catalyzed 1.V.S.R.. M. High yield bio-oil production from fast pyrolysis by metabolic controlling of Chlorella protothecoides. T. K. 1188-1195. 1330-1336.. 413-417. Supercritical Fluid Extraction of Nutraceuticals and Bioactive Compounds (pp. Y. Journal of Food Science. Kalva.3-diglyacylcerols by mixture response surface methodology... 89(12). T. S. (2008). 729-731. 513-524 McHugh. H. CO2 fixation and oil production Through microalga. (2008). Inoue. Ethanol production from CO2 by fermentative microalgae. and Hill. A. K. Fuel Processing Technology. Physical mechanism of ultrasound-assisted synthesis of biodiesel. and Kunito. (2006). (2009).. B. 279-288. D. Koegel. M. (2003). 343(1-2). A guide to the seaweed industry. (1995). F... V. Chowdhury. (2008).J. 9(7). R. J. K. J. Nurunnabi.Jr. W. Journal of Applied Phycology. 39-48. Tsai. Aryusuk. Okabe. and Wu. and Weirich. Ultrasonically driven continuous process for vegetable oil transesterification. B. Oil production from algal cells of dunaliella tertiolecta by direct thermochemical liquefaction. and Argyropoulos. (2008). Towards sustainable production of biofuels from microalgae. W. Mendes.. M. J. S.C.. Tran.. S. Lopez-Hernandez. T. and El Sheltawy.. Applied Biochemistry and Biotechnology. Y. Liao.T. (2007). C. A. B. Samejima. D. Yokoyama. (2007). 1735-1738.S. 14(4)..E. Mahajani. G. Fuel.. H. and Hoye.L. R. Catalysis Letters. Q. S. Applied Catalysis. International Journal of Molecular Sciences..N. 6. A continuous catalytic system for biodiesel production.. Energy Conversion and Management. A. Boyce.. C.T. Chang. Miao. J. D. Time factor in microwave-enhanced biodiesel production.. Vinatoru. Henrich. Pairintra. A: General. Stahl. Sivasankar. and Bandow. 128(1-2). Industrial & Engineering Chemistry Research. & Dumesic. (2008).F. and Shieh. and Okakura...C. (2004).. Murata. 250-254. Phillips. Nakanishi.. S. M. 36.. A.. (2008). R. (2005). Hon-Nami. Malik.T. and Naqiuddin. N.U. T. American Journal of Biochemistry and Biotechnology. 111(1-2). S. Yan. Refaat. Y.. Studies in Surface Science and Catalysis 114(Advances in Chemical Conversions for Mitigating CO2). 48(1). Technoeconomic analysis of a lignocellulosic biomass indirect gasification process to make ethanol via mixed alcohols synthesis.. Kishimoto. A unique feature of hydrogen recovery in endogenous starch-to-alcohol fermentation of the marine microalga. S. S.L. Minowa. 15(6). E. K.K. McNeff. Maeda. K. Biotechnology Letters.. and Yokoyama. Fischer-Tropsch Synthesis over Ru catalysts by using syngas derived from woody biomass..-Y.. 534-544. In Martinez. Garcia. 8887-8897.. McDermid. C. H. J. Microwave assisted in continuous biodiesel production from waste frying palm oil and its performance in a 100 kW diesel generator.. 66-71. An overview of aqueous-phase catalytic processes for production of hydrogen and alkanes in a biorefinery. Hirano. Hon-Nami. (1995). Hossain. Rasmussen. R. R.Q.. 46(26). K. Ogi. 119–132. K. Salleh. R.S. Nutritional composition of edible Hawaiian seaweeds. D. Fedie. Supercritical fluid extraction of active compounds from algae. Synthesis using bio-syngas and CO2. Algal Biofuel Conversion Technologies   59 . Energy for Sustainable Development. Catalysis Today. K.... (2007). 1857-1861.. M.R.. 129(1-3). X. (2003). 70(6)... 560-565. Taylor and Francis..

& Wu.. Xiang. F. and catalyst in the transesterification of soybean oil.. R. Yang.C. Synthesis of biodiesel using microwave absorption catalysts. Alarcon. Yung. Y.Y. 558-563. (2008).V. Energy and Fuels. Energy and Fuels. I. Y. Synthesis of biodiesel from mixed feedstocks and longer chain alcohols using an acid-catalyzed method. B. X. 55(3-4). Journal of Biotechnology.. glycerol.. and Wei.. J. J. Y. H.. and Clark.. Yang. I. 106(1-4). G. K. Wang. Annals of the New York Academy of Sciences.D. (2008). Corazza. 793-801. J. and Boocock.. and Adlercreutz. Zhong.. Jablonski.. L.. Energy and Fuels. Chinese Science Bulletin..A. Journal of the American Oil Chemists’ Society. (2008). 118-125. Biomass and Bioenergy. M. E. 1047-1052. and Zhu. Wahlen. 171-179. D. X. Influences of bacterial degradation on gases generated by pyrolysis of heterotrophically yellowing algae. Q. G.. I. Catalysis Today. Sheng. 110(11). A highly active and stable Fe-Mn catalyst for slurry Fischer-Tropsch synthesis. Grompone. 23(4). X. 10(7). Alckmin. Evaluation of marine algae as a source of biogas in a two-stage anaerobic reactor system... Applied Biochemistry and Biotechnology.. and Magrini-Bair. Iwasaki Y.M. (2005). B. N. 2805-2809. Kusdiana. 170-175. 548-552. J. P.. and Saka. Zhang. H.. Borges. G. W. Wu. J. C. L.. I.A. Vieitez. 32(4). G. Zhou. B. Oliveira.C.. Oliveira. I. and Fu. and Doisaki. (2008). Energy and Fuels. M. 23(1). W. Xu. Algal Biofuel Conversion Technologies   . Green Chemistry. Vergara-Fernandez.R. Corazza. Da Silva.. Yang.. (2009). B. Identification of triacylglycerols in the enzymatic transesterification of rapeseed and butter oil.. H.. and Jachmanian. Review of catalytic conditioning of biomass-derived syngas. Phase distributions of alcohol... Improvement in lipase-catalyzed methanolysis of triacylglycerols for biodiesel production using a solvent engineering method. Energy and Fuels.M. (2009). 1874-1887. 39(23). Miao. 83(12). Y. M. and Seefeldt..R. (2008). 2018-2022. G.. Su. Barney. Journal of Molecular Catalysis B: Enzymatic.. Vieitez.H. Preparation of mesoporous polyoxometalate-tantalum pentoxide composite catalyst and its application for biodiesel production by esterification and transesterification. Yamane T. L.Svensson.... 126.S. Effect of temperature on the continuous synthesis of soybean esters under supercritical ethanol. Yu. (2004). European Journal of Lipid Science and Technology. A. D.Y. S. D. Continuous production of soybean biodiesel in supercritical ethanol-water mixtures.A. Warabi.C. Xu. B. F. 338-344. and Velasco. L. 22(6). B.... 115(1-3). Roxana R. Zhang. (2006).M... Da Silva. 23(1). 22(4). Shimidzu N. Yuan. C.. Biodiesel fuel from vegetable oil by various supercritical alcohols. Borges. Q. 499-507.. Grompone. High quality biodiesel production from a microalga Chlorella protothecoides by heterotrophic growth in fermenters.. (1998). G.. A. Multiple intensified performance of an enzyme-catalyzed reaction in organic medium. Vargas.V. (2006). 864(Enzyme Engineering XIV). 60 6. and Li. N. 1007-1013. (1994). 746-755. 4223-4228. Guo.. (2008). and Jachmanian. Y. (2008).

primarily by the derived from microalgae and cyanobacteria. but considerably under a dollar for a commodity treatment (for nutrient removal) to provide additional producer. For the purposes of the Roadmap. a reasonably benefits to the technology.1). the production a biorefinery for utilization of every component of the of industrial. and nitrogen-containing components enhance the economics of the process. corresponding markets in the long term. either through Microalgae and Cyanobacteria chemical. as a “high value” product biorefinery the possibility of coupling biomass cultivation to a fine chemical producer might be well over several with CO2 mitigation (for carbon credits) and wastewater dollars/lb. valued chemical is defined as one that will cost roughly products. Vorlop. dyes. As summarized economics of the system. all primary components of algal biomass – carbohydrates. to be developed to handle the large amount of waste that biodegradable polymers. Exhibit 7. high-value and high-volume chemicals from biomass raw material must be considered as a means to amino acids. proteins and a variety of inorganic and complex organic molecules – 7. Moreover. 2004). Inorganics Proteins enzymes. $0. aromatic essences. Heat.00/lb. glycerol. reductant.1 An overview of the biorefinery concept Biorefinery Aquaculture Primary Refinery Industry Conversion Carbohydrates Raw biomass.30 . and they may vary from region to in Exhibit 7. 2007 7.2. and efficiencies and reduced environmental impacts may have a variety of specialty products such as bioflocculants.7. The topic of polysaccharides. 2007) address these options and discuss how some of them and must be considered in determining the economics of are better opportunities as they will not readily saturate the process. generation will inevitably compete for carbon.$1. cosmetics. flavors. electricity Energy Modified from Kamm and Kamm. This chapter will of algal biomass becomes feasible (Mooibroek et al. is predicted to be generated by the process. pharmaceuticals. poly-unsaturated fatty acids. Co-products   61 .. the concept of Under the biorefinery concept (Exhibit 7. enzymatic or microbial conversion (see Chapter 6).500 x106 lbs/yr. the generation of other been discussed (see Chapter 6). and can be produced at a volume of roughly 100 . novel technologies with increased coloring substances. fats (oils). and energy from photosynthesis. CO2 Products and Complex Fats residues from Substances conversion Special substances: processes pigments. these include products for human and animal region according to the cost of the raw material (Willke and nutrition. without invoking competing co. etc. anti-oxidants.1 Commercial Products from can be converted into different products. The nature of the end products and of the technologies A large number of different commercial products have been to be employed will be determined. hormones. Indeed. Using appropriate technologies. fertilizers and soil conditioners. this chapter will focus on co-products must be correspondingly low since their the non-fuel co-products. wastewater. Co-products The “guiding truth” is that if biofuel production is conversion of algal biomass to other biofuels has already considered to be the primary goal. and stable isotopes for research purposes. The use of terms such as “high volume” or “high value” This chapter will also address within the context of the can be extremely subjective.

000 t/yr Chlorella. 62 7. DHA by Crypthecodinium • Human Health Food Supplement: The consumption of and Schizochytrium. is expected to grow in the future. All other PUFAs to a lesser extent. which may be added to human food and production at volumes and pricepoints that meet consumer animal feed for their health promoting properties demands or 2) new co-products that are low cost and have (Benemann. is anticipated to continue until 1) a sufficient Polyunsaturated Fatty Acids (PUFAs) number of the challenges outlined earlier in the Roadmap for biofuel production have been overcome and associated Microalgae can also be cultured for their high content life cycle costs are reduced to realize sustainable biofuel in PUFAs.e. animal feed are Spirulina. et al. 2006). poultry. However. which is sold either fish (Benemann. 600 t/yr Nostoc. 1990. nascent algal-derived biofuels market: in the long term. 1986. the resulting non-oil algal improve the economic viability of some algae processes biomass is very likely to be unsuitable for use as an in the short-term.) and scales (sizes and numbers biomass up to a level of 5 . Isochrysis. significant amounts of lead. supplant the fossil fuels market) must be commodities • Animal Feed Additive: Microalgal biomass has also based to meet required volumes at price points acceptable been used with good results (i. 1. Chlorella. Scenedesmus. The market for microalgal animal feeds. arsenic. etc. these existing markets (and associated Spirulina. from 300 to 3. 1996).) as market for microalgal biomass for animal nutrition and a feed additive for cows. 2.. estimated Therein lies a major conundrum associated with the to be about 300 million US$. With the possible exception of the existing response. Malcolm et al. 2006). docohexaenoic acid (DHA). ca. Yet the existing fossil fuels market is and the aquaculture feeds. the DHA oil market is quickly growing. and far on a commercial scale by microalgae. ca. GLA from evening primrose oil). particularly given the fact that algae transportation fuels below their market price. US$. product. fertility. is expected to expand significantly. currently at about 2.200 t/yr Dunaliella. only DHA has been produced thus Spirulina (Arthospira).g. is quickly growing. Dunalliella.. commonly considered PUFAs are arachidonic acid (AA). have also been produced by microalgae (Borowtizka. increasing fuel supplies without drastically increasing price. GLA by Arthrospira. Co-products   . The Dunaliella. γ-linolenic acid (GLA).. Chlorella. etc. pigs. currently at ~700 future algal-based biofuels market (designed in part to million US$. gas from coal-fired power plants that will be used co-products of higher value in the marketplace must be to supply carbon dioxide to the cultures will contain pursued in order to offset the costs of production of algal. the goal of the industry is to produce animal feed. 500 t/yr Aphanizomenon. Nitzschia. 1999). The market size. 1990. the biofuels markets will involve volumes even dogs and cats. Phaeodactylum and Nitzschia (Spolaore is currently restricted to only a few species. Although it is clear that co-products may other toxic elements.000 t/yr Spirulina. 1990. sold for the health food market.10% (wt. AA has been shown to be Food and Feed synthesized by Porphyridium.5 billion A number of anti-oxidants. better immune to the consumer. 280 million US$.5 billion US$. it is important to note that since the flue massive lipid production dominates.By definition.) can be safely of commercial plants) that are significantly less than those used as a partial replacement for conventional proteins associated with the existing high-value algae-derived (Spoalore et al. The production includes ca. ca. 1994 and 1996). having presently a retail value of 1. cadmium and derived biofuels. weight gain. Spolaore et (e. Radmer. This situation. and soil fertilizer. 2004. market size for β–carotene is estimated to be greater than Pavlova. protein content and the level of unsaturated fatty value products or co-products from algae. horses. and eicosapentaenoic acid (EPA). 2006). Skeletonema. appearance.. e. and Thalassiosira. yet in the short term. Phaeodactylum.g. thereby are known to be effective at metal absorption. Radmer. 3. and is β–carotene from Dunaliella salina.. not commodity acids determine the nutritional value of microalgal products. Nostoc and Aphanizomenon are more cost-effectively produced from non-algal sources (Radmer. 2006). The most very large potential markets are developed.. microalgal (of biomass. and EPA by microalgal biomass as a human health food supplement Nannochloropsis. Pulz and Gross. 1996. al. The most prominent aquaculture of mollusks.000 US$ per kg (Spolaore et al.. Anti-Oxidants The market. and ca. Both the production plants and distribution channels) are for high. The main species used in products. • Aquaculture: Microalgae are also used as feed in the Benemann. Chlorella and Scenesdesmus. However. Most as an extract or as a whole cell powder ranging in price frequently used species are Chaetoceros.. Although small. In poultry rations. crustaceans (shrimp). Tetraselmis. Nannochloropsis.

. growth promoters.. has been successfully used as a salmon feed to cosmetics. have also been used in the feed of carp and even chicken (Puls and Microalgae-produced coloring agents are used as natural Gross.500 Pulz&Gross (2004) Pulz&Gross (2004) Aquaculture 1.e. no current commercial market Spolaore et al. Co-products   63 .200 >280 Spolaore et al. 1986. Phycobiliproteins. (2006) Animal Feed Additive No available information 300 Pulz&Gross (2004) POLY-UNSATURATED FATTY ACIDS (PUFAs) ARA No available information 20 Pulz&Gross (2004) Pulz&Gross (2004) DHA <300 1.Coloring Agents the related carotenoids lutein and zeaxantin. and as fluorescent reagents in clinical or research give the fish meat a pink color preferred by the consumers laboratories (Spolaore et al. and research. Spolarore et al.. 2006). the cyanobacterium Arthrospira and the rhodophyte Astaxanthin. dyes for food.000 soil conditioners Metting&Pyne (1986) 7... are used as food dyes. or as pigments i.. (2006) Phycocyanin No available information >10 Pulz&Gross (2004) Phycoerythrin No available information >2 Pulz&Gross (2004) FERTILIZERS/SOIL CONDITIONERS Fertilizers. (2006) EPA Potential product. produced by in animal feed (Borowitzka.. and Exhibit 7. 2006). Pulz&Gross (2004) No available information 5.000 2. 2003. a carotenoid produced by Hematococcus Porphyridium..000 700 Spolaore et al. 2004. phycoerythrin and phycocyanin. 1990). (2006) PUFA Extracts No available information 10 Pulz&Gross (2004) GLA Potential product. (2006) ANTI-OXIDANTS Pulz&Gross (2004) Beta-Carotene 1. (Olaizola. Spolaore et al. (2006) Tocopherol CO2 Extract No available information 100-150 Pulz&Gross (2004) COLORING SUBSTANCES Pulz&Gross (2004) Astaxanthin < 300 (biomass) < 150 Spolaore et al..500 Spolaore et al. no current commercial market Spolaore et al. pigments in pluvialis. 2006).. Astaxanthin. Benemann. cosmetics.2 Summary of microalgae commercial products market MARKET SIZE SALES VOLUME COMMERCIAL PRODUCT REFERENCE (TONS/YR) (MILLION $US/YR) BIOMASS Health Food 7.

Pulz and Gross.. 2006)..2 Commercial Products factor. polysaccharides (Benemann. cost of the new product must be equivalent to the 1986.3). biopolymers and biodegradable plastics (Philip et al. The market for these specialty products is likely to nutritional products. Microalgal biomass could in principle serve the same purpose. 1986). Alginate. pharmaceutical. The production bioactive compounds (Burja et al. be very small due to their specialized applications.3 Potential Options for the derived from microalgae (Metting and Pyne. material it replaces and to be competitive typically. 2001. the only issue is price. Identical to an existing chemical. fuel or other product. Identical in functional performance to an existing chemical. 1990. Exhibit 7. 1990).. or with replacements such as algal polysaccharides that are extracted for their commercial proteins for distillers dried grains from corn dry grind value (Exhibit 7. Here price is a major 7. Olaizola. plant growth regulators could be 7. Recovery of Co-products Other Specialty Products Co-products from algal refineries should address one There are a number of specialty products and chemicals of these three criteria to be commercially viable and that can be obtained from microalgae. Pulz and Gross. These include acceptable: bioflocculants (Borowitzka. 2003.3 Global value of seaweed products per annum (McHugh. and stable isotopes for material (shutdown economics). Co-products   . or other product... pharmaceuticals and In this instance. pet food. Metting and Pyne. Singh et al.e. Price becomes less of an issue if algae and agar and carrageenen from red algae. 2001). macroalgae (i. toothpaste) $240 million Other uses of seaweeds Fertilizers and conditioners $5 million Animal Feed $5 million Macroalgal Biofuels Negligible TOTAL $5. 2004). etc. 1990. seaweeds) are used as a plant the redox balance favors formation of pyruvate as the end fertilizer and to improve the water-binding capacity and product (Forro. pharmaceutical. mineral composition of depleted soils (Metting et al. cosmetics (Spolaore et al. the product can be labeled “organic” and thus saleable at a premium. kombu. food additive. Borowitzka. They include alginate from brown ethanol processing.Fertilizers which occurs in high concentrations in brown seaweeds. Pulz and Gross. but the source of the material can often provide from Macroalgae some advantage. fuel. 2.5-6 BILLION 64 7. 2007. Radmer. Furthermore. is considered recalcitrant to ethanol fermentation since Currently. 1986). 1987).. 1994. medical) $213 million Carrageenen (Food additive. it 2004). 2003) PRODUCT VALUE Human Food (Nori. 2005. must be produced at a cost 30% lower than the existing 1986.) $5 billion Algal hydrocolloids Agar (Food ingredient.. wakame. Wu et al. but has been achieved for some chemicals and proteins/ 2004). This occurs with natural oils which manufacturers in many cases would prefer if the costs Macroalgae possess high levels of structural were comparable. aonori. 1. This sets a high bar research (Benemann. biological/microbiological) $132 million Alginate (Textile printing.

. and the glycerol as nutrients for algal cultivation or could be sold as soil from the transesterification of lipids to biodiesel fertilizers and conditioners.4 Overview of the five Algal Biomass potential options for the recovery and use of co-products Option 1 Extract Lipids Energy for Fuel Burn Residue Ash/ Soil Amendments Option 5 Soil Fertilizer/ Conditioners Dry Residue Processing Proteins as Option 2 Co-Products Food/ Feed Supplement Processing Option 3 Non-Fuel Surfactants/ Bioplastics Lipids Chemical/ Biological Conversion Option 4 Carbohydrates Ethanol/ Butanol/ Glycerol Chemical/ Biological Conversion 7. such as pyrolysis. 2009). is the anaerobic • Option 1 – Maximum energy recovery from digestion of lipid-extracted biomass.pdf).3 l per gram volatile solids (Huesemann • Option 2 – Recovery of protein from the lipid.4). Residuals as Soil Amendments these technologies are still in the testing and development Given the large amounts of lipid-extracted biomass residues stage. These technologies are able to convert Lipid-Extracted Biomass. The economic value of the produced extracted biomass for use in food and feed methane is equivalent to about $100 per ton of digested • Option 3 – Recovery and utilization of non-fuel lipids biomass. Maximum Energy Recovery from the (see Chapter 6). as soil fertilizer and conditioner Each option and its associated technologies and future In addition to anaerobic digestion. Co-products   65 . which could then be either sold on the new product. the issues enough markets for potential co-products. with reported methane yields of about 0. could also be potentially considered for the recovery of energy from the lipid-extracted biomass Option 1. with potential whole (i. The residuals • Option 4 – Recovery and utilization of carbohydrates remaining after anaerobic digestion could either be recycled from lipid-extracted biomass. and because of their large energy inputs (temperature that will likely be generated in future microalgal biofuels and pressure). it may be difficult to identify large performance characteristics. There are at least five different options for recovering economic value from the lipid-extracted microalgal biomass The most promising energy recovery technology. Recycle Nitrogen Gases Exhibit 7. could have poor or even negative energy balances (Huesemann and Benemann. and Benemann. Therefore. the open market or used on-site in the various biorefinery operations. one are less related to costs and more to the functional option would be to convert as much of the lipid-extracted performance and potentially enhanced performance of biomass into energy. thermochemical research needs are discussed below. Nevertheless. non-extracted) micro and macroalgal biomass use of residuals as soil amendments has been successfully demonstrated.3. and combustion.unh. with use of the residual biomass edu/p2/biodiesel/pdf/algae_salton_sea. lipids only. However. 2009). These are: from a practical and economic perspective.e. as is currently already done for • Option 5 – Recovery/extraction of fuel certain waste water treatment sludges (see http://www. both (Exhibit 7. In this case. conversion technologies. Anaerobic digestion of the lipid extracted biomass. with Potential Use of a much larger fraction of biomass into fuels. gasification. New material with unique and useful functional production systems. which is significant in terms of reducing the overall cost of liquid biofuels production.

current levels.. Choi. succinic. Kamm and Kamm.usda. detergents. 2007). also benefit from supplementation with microalgal proteins. and the Glycerol from the to approximately 60 million tons per year for distillers dry Transesterification of Lipids to Biodiesel grains plus soluble (DDGS)) (Berger and Good. 2009. 2007. and (c) at products such as polymers. it may be possible to recover important industrial production of green solvents. green solvents and a result. 2006). poultry. acetic. human nutrition may organic acids (formic. Option 2.the thermochemical conversion of lipid-extracted biomass Option 3. is potentially very large.e. could also be anaerobically pretreating lignocellulosic feedstocks prior to fermentation fermented to the above mentioned and other end products to fuel ethanol. this option (Datta et al. generate electricity directly in biofuel cells (Yildiz and 66 7. pigs. the market for animal feed (cattle.g. (Hu et al. glycerol could be used to potentially very large. Hydrogen and The byproduct material. As of industrial surfactants.. i. and paint (Yazdani and Gonzalez. Kawaguchi et al. while butanol and organic make a useful animal feed. 2007).20 per gallon ethanol. 1 billion kg. thereby depending on the algal species and culturing conditions reducing the expense for nitrogen fertilizers.. the individual local markets for animal feed are often not succinate is an intermediate in the production of a variety located adjacent to areas where algae may be produced. propionic. As was pointed out above. antifreeze. For example. ethanol.. (b) approximately 100 grams which is used in the formulation of a variety of industrial of cellulase are needed per gallon of ethanol. However. than $0. epoxies. and as a renewable substitute (Zerlov et al. product into the appropriate range or rearrange the carbon chains through catalytic cracking and catalytic reforming.. The current price for DDGS ranges from $110 . Recovery and Utilization of Carbohydrates pets) is already very large and growing (estimated to rise from Lipid-Extracted Biomass. Recovery and Utilization of Non-fuel Lipids has the potential advantage that the resulting nitrogen- It is well known that microalgae can synthesize a variety containing gases (e. feeding studies acids could serve as renewable feedstocks for the chemicals indicate that algae cannot be used as a high percentage industry. ammonia and nitrous oxides) could of fatty acids with carbon numbers ranging from C10 to C24. biodegradable plastics (Kamm and Kamm. byproduct markets are likely to be overwhelmed. adhesives. least 10. the projected market for cellulases is 2007. would require the use of specially selected or engineered microalgal strains capable of producing these enzymes. be recycled into the microalgal culture ponds. etc. the residual microalgal (http://www. relative derived ethylene and propylene. due to issues such as taste of the meat or eggs. and lactic) (Huesemann and Benemann. which contains proteins. Recovery of Protein from the Lipid-Extracted It may be worthwhile. Furthermore.. Similarly. and Option 4. 2007). and its applications enzymes such as cellulases or other industrial enzymes include the pharmaceutical and agrochemical industries from the lipid-extracted biomass. butyric. to separate specific Biomass for Use in Food and Feed lipids present in the algal oil that have utility as chemical Following the extraction of lipids from the microalgal feedstocks for the manufacture of surfactants. However.5 billion gallons of lignocellulosic ethanol will be solvents. fish. The Glycerol. Furthermore. 2008). thus butanol could serve to the amount of byproduct that would be produced. glycerol microalgal cellulases could be provided at a cost of less could be converted by certain bacteria to 1. specifically cellulases for microalgal lipids to biodiesel. a food and feed supplement.3-propanediol. produced by 2020. for chemical synthesis of a variety of products. the polymers that are currently produced from fossil oil- overall size of the animal feed market is small. 2008). polycarbonates and polyurethanes. and could be advantageous. Furthermore. is the starting material for the production of polyester. Furthermore. 2001).txt). Co-products   . it is also used in the In addition. and butanol). a byproduct of the transesterification of market for industrial enzymes. Since biomass may contain sufficient levels of carbohydrates that protein is generally the key and often limiting ingredient could be converted through anaerobic dark fermentations in animal feed. Assuming that (a) (Yazdani and Gonzalez. Since the generation of gasoline.150 per ton After the extraction of lipids. might ethanol could be used as biofuel. it will be necessary to either separate the soil amendment. however.ams. including and interactions with animal digestion. the protein fraction and specialty products such as urethanes. from the residual biomass could be extracted and used as lubricants. jet fuel. 1995). Finally. Lactic and byproduct prices will be greatly depressed versus acid. the mineral-rich ash generated by these thermochemical and diesel substitutes will require specific ranges of carbon processes could possibly be used for nutrient recycle or as a chain length. solvents (acetone. aliphatic polyesters. butanol is a valuable C4 compound of feed. biomass for liquid biofuel production. which can be converted into polypropylene oxide. supplementation with microalgal proteins to hydrogen.

W. Technological and economic potential of poly(lactic acid) and lactic acid derivatives. Crouch. J. (91). J.. M.A. H2 production from algal biomass by a mixed culture of rhodobium marinum A-501 and lactobacillus amylovorus. (1990). B. (16). B.. J. Metting. Biologically active compounds from microalgae. polysaccharides for use as emulsifiers in food and industrial non-fuel lipids. 9347-9377. J. 589-627. Burja. (2003). Developments in Aquaculture and Fisheries Science (Netherlands).C. Metting. (105). it applications (Mooibroek et al. Seibert. and P. Q. D. J. The alga dunaliella: biodiversity. Benemann. Advances in Biochemical Engineering/Biotechnology. (8). Borowitzka. (2007). Wright. and Conditioner In case none of the above mentioned four options are It may also be possible to extract microalgal economical. (2006).S. Enzyme Microbial Technology. if desired. and carbohydrates is not cost-effective.M. K. Tsai. E. J. Champaign-Urbana. organic matter as soil fertilizer and conditioners. and potential. Microalgal triacylglycerols as feedstocks for biofuel production: Perspectives and advances. R&D efforts as discussed under Chapter 2. Posewitz. Ghirardi. Moon. Algal Biology. Food and Agriculture Organization.. and the development of genetic engineering the formation of more than 400. SPB Academic Publishing. W. University of Illinois. carbohydrates. (2008). this option will also require and potentially growing..jsp?article_id=377). 3(12).. (2001). Burgess. L. M. Suppl. FEMS Microbiological Review. (1986). with saturated. Banaigs. Bonsignore. the recovery and use of energy.G. (1987). Introduction to Applied Phycology. with special focus on Dunaliella sp. (57). the issue of scale enters in. (2001). genomics. Marine cyanobacteria – A prolific source of natural products. Sommerfeld. New Hampshire. Biorefineries – multi product processes. Microbial degradation of marine biomass. 173-180. (1990). proteins. (54). Good. Recent Patents on Biotechnology. these are the development of high throughput technologies for the quantitative characterization of References Benemann. S. B. specifically. Darzins.J. S. M. (31). 277-282. 247-256. Choi. McHugh.. Huesemann... Distillers dried grains plus solubles utilization by livestock and poultry in corn-based ethanol in Illinois and the U. Miyamoto. M. (2007).L. K. As the current production levels for biodiesel (700 million gallons in 2008) already has the market for glycerol Option 5. Tetrahedon. I. K. 7. 621-63. the market for organic fertilizer is large As was the case with Option 3. van Staden. Abou-Mansour. microalgal metabolites. E. carbohydrates could be recycled into pulp and paper which involves the extraction of only fuel lipids and the streams. The Plant Journal..P. As was mentioned above. Kamm. substituting for lignocellulosic materials derived subsequent use of the biomass residues rich in nitrogen and from forestry resources. M. A guide to the seaweed industry. Finally. Frank. Glycerol-based biorefinery for fuels and chemicals. 386-394. M.J. J. (2). Jarvis. Microbiological Sciences. Berger. Microalgae products and production: An Forro. Netherlands. 1994). R. and biotechnology. D. processes. 5. Zimmerman. Hashimoto. (2008). Biofuels from microalgae: Review of products. Once again. additional capacity from algal lipids may find it Use of the Residual Biomass as Soil Fertilizer exceedingly difficult to find uses. A Report from the Department of Agricultural and Consumer Economics. L.H. Hirata. including sugars and complex Production of 1 billion gallons of biodiesel will result in carbohydrates. Microalgae as sources of fine chemicals.R. Journal of Industrial Microbiology. Developments in industrial microbiology.000 tons of glycerol (http:// tools to improve yields of desired products. 2007). M. microalgal is possible to revert to the most simple option (Option 5). Kamm.H. Pyne. A.e. (2009).R. No. W. P. B. Hu. i.Kadirgan. 221-231.175-204. Agronomic uses of seaweed and microalgae. J. A.biodieselmagazine. Co-products   67 . including www.R. Kawaguchi H. Datta. 372-375. Science Publishers. physiology. The Hague. (1986). Journal of Biosciences Bioengineering. Recovery (Extraction) of Fuel Lipids Only.

(2007).Y.Mooibroek. Industrial bioconversion of renewable resources as an alternative to conventional chemistry.N. Applied Microbiology Biotechnology. Zerlov. W. S. Current Opinion Biotechnology. (2004).. G. (1994).H. Oosterhuis. 233-247. (141). O. T. (2004). Biomolecular Engineering. E. Applied Microbiology and Biotechnology. 87-96. (2007). (6). V. T. Electrocatalytic oxidation of glycerol.Y. (82). Velikodvorskaya. Kadirgan. 213-219. S. Duran..Valuable products from biotechnology of microalgae.. Journal of Chemical Technology and Biotechnology. A. 73-95..J.C. A. G. Kate. (18). W. Shen. Isambert.H. R. Commercial applications of algae: Opportunities and Constraints. 263-270. Journal of Applied Phycology. Spolaore.. M. Keshavarz. (2007). R. I. Olaizola. Bioresource Technology. (77). H. Anaerobic fermentation of glycerol: A path to economic liability for the biofuels industry. (2006). 85-90. Commercial applications of microalgae. Singh. S. (20). Sanders.A. Polyhydroxyalkanoates: Biodegradable polymers with a range of applications. Wu. (2006). Steinbüchel. (25). Journal of Electrochemical Society. Radmer. (2005). 257-267. Berezina. Vorlop. Franssen. Joannis-Cassan. 101(2). (1996). 459-466. Commercial development of microalgal biotechnology: From the test tube to the marketplace. 635-648. Z.J. Scott. M. Accumulation of poly-b-hydroxybutyrate in cyanobacterium synechocystis sp. 46(4).. (65). (71). Yazdani. R.F. Philip. (66). 93-98. Radmer. N. Critical Reviews in Biotechnology. B.C. Parker.S. Applied Microbiology and Biotechnology.. G. K. (76)... C. Banerjee. 68 7.D. Co-products   . O. 587-597. (1994). Journal of Bioscience and Bioengineering. 723-730. (2003). Willke. B. J. Applied Microbiology and Biotechnology. Wu... Toonen. Q. Assessment of technological options and economical feasibility for cyanophycin biopolymer and high-value amino acid production. M. PCC6803. F. (2001). Bacterial acetone and butanol production by industrial fermentation in the Soviet Union: Use of hydrolyzed agricultural waste for biorefinery. Gross.V. Roy. Schwarz. Giuseppin. Algal diversity and commercial algal products. Bioactive compounds from cyanobacteria and microalgae: An overview. U. H. 131-142. Gonzalez. Yildiz. P. Pulz. BioScience. E.

products – ethanol. transporting. These have been challenging issues appear to be more compatible with the existing petroleum for lignocellulosic feedstocks (Hess et al. and final fuels biomass. and bioproducts.. high capacity. higher alcohols. and end-use vehicles). other distribution aspects pipelining. with some evidence suggesting scalability. rails. biofuels. It is also known that temperature and storage material have important • Maximizing the substrate energy-density and stability. However. and final fuels from the systems. poorly characterized. similar to plant lipids may be possibly taken into pipelines. few studies exist that While the demonstration flights mitigate some address outstanding issues of storing. to be compatible for use in select demonstration flights (Buckman and Backs. Thus.. as corrosivity) for trucks. and hydroreformed biofuels – are being considered harvesting conditions differing in salinity. remain poorly studied: Being intermediate steps in the supply chain. The optimal Technologies). biogas. As a variety of fuel rate of algae decomposition (Rieper-Kirchner. Ethanol. Distribution and Utilization  69 . the stability of the algal biomass under different decision points. it is difficult to predict how utilization challenges associated with each of these possible these factors will influence biomass storage and opportunities is discussed. production. conforming to existing standards (e. biofuels oils (Miyashita and Takagi. delivery vehicles. and jet fuels are all possible products for energy-inputs and costs. hazardous Distribution is complicated by the fact that several materials and associated human health impacts. Efstathiou and Credeur. depending on whether it will be dewatered/ • Optimizing the scale of operations to the parameters densified biomass and/or fuel intermediates that are to stated above. and the quality of the final fuel product. transport. renewable transport method(s) should be analyzed and optimized gasoline. and the ultimate economics and operations of that natural bacterial communities increase the an integrated algal biofuels refinery. container dimensions. such as cultivation and harvesting. blending. jet-fuel blends from a variety can be expected to influence the economics of algal of oil-rich feedstocks. within the context of from algal biomass. • Second. Each of these different fuels has where the algae production and biorefinery facilities different implications for distributions. fuel intermediates. fuel intermediates. and barges is critical discussed in detail in Chapter 6 (Algal Biofuel Conversion to minimizing infrastructure impacts. pH and from algal biomass resources.g. diesel. biodiesel.. including algae. 1986). fuels intermediates. It is conceivable that these “bio-crudes” Lowering costs associated with the delivery of raw will be compatible with current pipeline and tanker biomass. have been shown biofuels as well. Some of these fuels are to be sited. it is known that the presence feedstock production center to the ultimate consumer of unsaturated fatty acids causes auto-oxidation of are common goals for all biofuels. pyrolysis oil. biodiesel. combusting. infrastructure concerns. storage tanks. for ideal lipid compositions. 2008). be transported to the refinery. 2009.1 Distribution of algal lipid intermediates have not yet been established. implications for biodiesel stability (Bondioli et al. 8. and transport scenarios is these logistics through end-use issues influence siting. In turn. Distribution and Utilization Distribution and utilization are challenges associated with It is also anticipated that gasoline and diesel range fuels virtually all biofuels. In all cases. and completely compatible with the existing liquid transportation fuel infrastructure. Specifically. dispensing equipment.g.. 2009) and infrastructure. the specific distribution and dewatering levels. storage. account for the storage of algae lipids (Hu et al. which carries infrastructure costs can be lowered in four ways: implications for the producers of algae and selection • Minimizing transport distance between process units. • Maximizing compatibility with existing infrastructure 1995).8. and • Third. In the context of a variety of culturing and syngas. and different fuels from algae are being considered. and dispensing algal concerning algal biomass. an issue that impacts oleaginous microalgae feedstocks is that the transport and storage mechanisms 8. Although the biofuel product(s) from algae will not require significant distribution system from algal biomass would ideally be energy-dense modifications during or after processing in the refinery. 2009). 1990). materials and temperature considerations (e. distribution and utilization need to be discussed in the context of earlier • First.

Ethanol is another likely fuel from algae. With over Typically, compliance with specifications promulgated by
10 billion gallons per year produced and consumed organizations such as ASTM International ensures that a
domestically, distribution-related issues for ethanol has fuel is fit for purpose (ASTM International, 2009a, 2009b,
been studied for some time, and algal ethanol can benefit 2009c, 2009d, and 2009e). Failure of a fuel to comply with
from these analyses. While not as energy dense as purely even one of the many allowable property ranges within
petroleum-derived fuels, ethanol is an important fuel the prevailing specifications can lead to severe problems
oxygenate that can be used in regular passenger vehicles in the field. Some notable examples included: elastomer-
and special flex-fuel vehicles at up to 10% and 85% gasohol compatibility issues that led to fuel-system leaks when
blends, respectively. However, considerable infrastructure blending of ethanol with gasoline was initiated; cold-
investments need to be made for higher ethanol blends weather performance problems that crippled fleets when
to become even more attractive and widespread. One blending biodiesel with diesel was initiated in Minnesota
issue is that ethanol is not considered a fungible fuel; it in the winter; and prohibiting or limiting the use of the
can pick up excessive water associated with petroleum oxygenated gasoline additive MTBE in 25 states because
products in the pipeline and during storage, which causes it has contaminated drinking-water supplies (McCarthy
a phase separation when blended with gasoline (Wakeley and Tiemann, 2000). In addition to meeting fuel standard
et al., 2008). One possible way to address this is to build specifications, algal biofuels, as with all transportation
dedicated ethanol pipelines; however, at an estimated fuels, must meet Environmental Protection Agency
cost of $1 million/mile of pipeline, this approach is not regulations on combustion engine emissions.
generally considered to be economically viable (Reyold,
2000). Another possibility is to distribute ethanol blends by As is true of any new fuel, it is unlikely that new
rail, barge, and/or trucks. Trucking is currently the primary specifications will be developed for algal fuels in the near
mode to transport ethanol blends at an estimated rate of term (i.e., at least not until significant market penetration
$0.15/ton/kilometer (Morrow et al., 2006). This amount is has occurred); hence, producers of algal fuels should
a static number for low levels of ethanol in the blends (5% strive to meet prevailing petroleum-fuel specifications.
to 15%); as the ethanol content in the blend increases, the Nevertheless, research and technology advancements
transport costs will also increase due to the lower energy may one day yield optimized conversion processes which
density of the fuel. can deliver algae-derived compounds with improved
performance, handling, and environmental characteristics
8.2 Utilization relative to their petroleum-derived hydrocarbon
counterparts. If significant benefits can be demonstrated,
new specifications can be developed (e.g., ASTM D6751
The last remaining hurdle to creating a marketable new
and D7467).
fuel after it has been successfully delivered to the refueling
location is that the fuel must meet regulatory and customer
requirements. As mentioned in Chapter 6, such a fuel is The discussion below is divided into separate sections that
said to be “fit for purpose.” Many physical and chemical deal with algal blendstocks to replace gasoline-boiling-
properties are important in determining whether a fuel is fit range and middle-distillate-range petroleum products,
for purpose; some of these are energy density, oxidative and respectively. These classifications were selected because
biological stability, lubricity, cold-weather performance, the compounds comprising them are largely distinct and
elastomer compatibility, corrosivity, emissions (regulated non-overlapping. Within each of these classifications,
and unregulated), viscosity, distillation curve, ignition hydrocarbon compounds and oxygenated compounds are
quality, flash point, low-temperature heat release, metal treated separately, since their production processes and in-
content, odor/taste thresholds, water tolerance, specific use characteristics are generally different.
heat, latent heat, toxicity, environmental fate, and sulfur
and phosphorus content. Petroleum refiners have shown Algal Blendstocks to Replace
remarkable flexibility in producing fit-for-purpose fuels
Middle-Distillate Petroleum Products
from feedstocks ranging from light crude to heavy crude,
oil shales, tar sands, gasified coal, and chicken fat, and are Petroleum “middle distillates” are typically used to create
thus key stakeholders in reducing the uncertainty about the diesel and jet fuels. The primary algae-derived blendstocks
suitability of algal lipids and carbohydrates as a feedstock that are suitable for use in this product range are biodiesel
for fuel production. (oxygenated molecules) and renewable diesel (hydrocarbon
molecules). The known and anticipated end-use problem
areas for each are briefly surveyed below.

70 8. Distribution and Utilization 

Oxygenates: Biodiesel isomerized (i.e., transformed from straight- to branched-
chain paraffins), renewable diesel and SPK will have
Biodiesel is defined as “mono-alkyl esters of long
cold-weather performance problems comparable to those
chain fatty acids derived from vegetable oils or animal
experienced with biodiesel. Also, as was the case with algal
fats” (ASTM International, 2009b). Biodiesel has been
biodiesel, contaminants and end-product variability are
demonstrated to be a viable fuel for compression-ignition
engines, both when used as a blend with petroleum-derived
diesel and when used in its neat form (i.e., 100% esters)
(Graboski and McCormick, 1998). The primary end-use Algal Blendstocks for Alcohol and
issues for plant-derived biodiesel are: lower oxidative Gasoline-Range Petroleum Products
stability than petroleum diesel, higher emissions of nitrogen
oxides (NOx), and cold-weather performance problems While much of the attention paid to algae is focused on
(Knothe, 2007). The oxidative-stability and cold-weather producing lipids and the subsequent conversion of the lipids
performance issues of biodiesel preclude it from use as a to diesel-range blending components (discussed above),
jet fuel. The anticipated issues with algae-derived biodiesel algae are already capable of producing alcohol (ethanol)
are similar, with added potential difficulties including: directly, and there are several other potential gasoline-range
1) contamination of the esters with chlorophyll, metals, products that could be produced by algae-based technology/
toxins, or catalyst poisons (e.g., sulfur and phosphorus) biorefineries. Petroleum products in the alcohols and
from the algal biomass and/or growth medium; 2) undesired gasoline range provide the major volume of fuels used by
performance effects due to different chemical compositions; transportation vehicles and small combustion engines in
and 3) end-product variability. the United States. Ethanol or butanol are the most common
biofuels currently being considered for use in gasoline,
and these alcohols can be produced from fermentation of
Hydrocarbons: Renewable Diesel and starches and other carbohydrates contained in algae.
Synthetic Paraffinic Kerosene
The hydrocarbon analog to biodiesel is renewable diesel,
which is a non-oxygenated, paraffinic fuel produced Additionally, the hydro-treating of bio-derived fats or
by hydrotreating bio-derived fats or oils in a refinery oils in a refinery will typically yield a modest amount of
(Aatola et al., 2009). Algal lipids can be used to produce gasoline-boiling-range hydrocarbon molecules. Refiners
renewable diesel or synthetic paraffinic kerosene (SPK), refer to this material as “hydro-cracked naphtha.” This
a blendstock for jet fuel. These blendstocks do not have naphtha tends to have a very low blending octane and
oxidative-stability problems as severe as those of biodiesel, would normally be “reformed” in a catalytic reformer
and renewable diesel actually tends to decrease engine- within the refinery to increase its blending octane value
out NOx emissions. Nevertheless, unless they are heavily prior to use in a gasoline blend.

8. Distribution and Utilization  71

Aatola, H., Larmi, M., Sarjovaara, T., and Mikkonen, S. (2009). Hydrotreated vegetable oil (HVO) as a renewable diesel fuel: Trade-off
between NOx particulate emission and fuel consumption of a heavy duty engine. SAE International Journal of Engines. 1(1), 1251-
ASTM International. (2009a). Standard specification for diesel fuel oils. West Conshohocken, PA: ASTM International. Available online
ASTM International. (2009b). Standard specification for biodiesel fuel blend stock (B100) for middle distillate fuels. West Conshohocken,
PA: ASTM International. Available online at
ASTM International. (2009c). Standard specification for aviation turbine fuels. West Conshohocken, PA: ASTM International. Available
online at
ASTM International. (2009d). Standard Specification for Automotive Spark-Ignition Engine Fuel. West Conshohocken, PA: ASTM
International. Available online at
ASTM International. (2009e). Standard specification for diesel fuel oil, biodiesel blend (B6 to B20). West Conshohocken, PA: ASTM
International. Available online at
Bondioli, P., Gasparoli, A., Lanzani, A., Fedeli, E., Veronese, S., and Sala, M. (1995). Storage stability of biodiesel. Journal of the
American Oil Chemists’ Society.72(6), 699-702.
Buckman, R. (2009). Boeing backs biofuels. Retrieved October 26, 2009, from at
Efstathiou, J. and Credeur, M.J. (2009). Algae-powered jet proves biofuel in continental test (update 2). Retrieved October, 23, 2009 from at
Frega, N., Mozzon, M. and Lercker, G. (1999). Effects of free fatty acids on oxidative stability of vegetable oil. Journal of the American
Oil Chemists’ Society. 76(3), 325-329.
Graboski, M.S. and McCormick, R.L. (1998). Combustion of fat and vegetable oil derived Fuels in diesel engines. Progress in Energy and
Combustion Science. 24(2), 125-164.
Hess, J., Kenney, K., Wright, C., Perlack, R., and Turhollow, A. (2009). Corn stover availability for biomass conversion: Situation analysis.
Cellulose. 16(4), 599-619.
Hu, Q., Sommerfeld, M., Jarvis, E., Ghirardi, M., Posewitz, M., Seibert, M., and Darzins, A. Microalgal triacylglycerols as feedstocks for
biofuel production: Perspectives and advances. The Plant Journal. 54(4), 621-639.
Knothe, G. (2007). Some aspects of biodiesel oxidative stability. Fuel Processing Technology. 88(7), 669-677.
McCarthy, J.E. and Tiemann, M. (2000). MTBE in gasoline: Clean air and drinking water issues. Washington, D.C.: Congressional
Research Service, The Library of Congress. Available online at
Miyashita, K. and Takagi, T. (1986). Study on the oxidative rate and prooxidant activity of free fatty acids. Journal of the American Oil
Chemists’ Society. 63(10), 1380-1384.
Morrow, W.R., Griffin, W.M., and Matthews, H.S. (2006). Modeling switchgrass derived cellulosic ethanol distribution in the United
States. Environmental Science & Technology. 40(9), 2877-2886.
Reyold, R. (2000). The current fuel ethanol industry transportation, marketing, distribution and technical considerations. Bremen, IN:
Downstream Alternatives, Inc. Available online at
Rieper-Kirchner, M. (1990). Macroalgal decomposition: Laboratory studies with particular regard to microorganisms and meiofauna.
Helgoland Marine Research. 44(3), 397-410.
Wakeley, H.L., Griffin, W.M., Hendrickson, C., and Matthews, H.S. (2008). Alternative transportation fuels: Distribution infrastructure for
hydrogen and ethanol in Iowa. Journal of Infrastructure Systems. 14(3), 262-271.

72 8. Distribution and Utilization 

off-shore marine environment CO2 sources with algae production are outlined. Integration with wastewater variety of conditions may be more or less cost-effective treatment can play an additional important role in the for the siting and operation of algal biomass production. Maxwell et al. This chapter provides an overview of the issues associated with site location and key resources for various microalgae Exhibit 9. This scenario Exhibit 9. availability. water.g. needs to be analyzed from a site location. These parameters with wastewater treatment and industrial sources of are of greatest importance to siting. microalgae to significantly contribute to meeting U. labor. For each parameter. and other inputs required for siting and operating industrial Heterotrophic microalgae production bioreactor-based algae production and post-processing to fuels and other co-products Availability of suitable coastal and off-shore marine site loca- Photoautotrophic macroalgae production tions 9. Sufficient land. energy. supplemental CO2 supply.2 provides a simple high-level overview of and macroalgae production approaches (Exhibit 9. characteristics. energy.1 Resource Requirements for production. 1990. a for algal biofuel production. and conditions of the particular algae species being cultivated CO2 resources were identified at the time to suggest and the engineered growth systems being developed and that the production of billions of gallons of algal deployed. land-based photoautotrophic microalgal terms of their geo-location. an in-depth discussion is included on the potential pertain to the algae biofuels production inputs of climate. Similarly. as with any biomass-based technology and industry. In addition to coastal and inland photoautotrophic the challenges associated with coupling industrial microalgae production. 1990). concentrated CO2. Critical requirements—such as suitable land and climate. 1998). and land. Photoautotrophic Microalgae Approach sustainable water resources. water. CO2.S. and from possible nutrient recycling from residual and other inputs associated with facilities infrastructure algal biomass.9. CO2. transportation fuel needs and displacing petroleum use.1 Key resource issues for different algae systems ALGAE PRODUCTION APPROACH KEY RESOURCE REQUIREMENTS Climate. Resources and Siting The development and scale-up of algal biofuels 9. To achieve success regarding both Program (Sheehan et al. sourcing of nutrients from both the input wastewater Additional resources include materials. The status of algae-based wastewater and conducting operations and maintenance.. as Different Cultivation Approaches well as from a resource availability and use perspective. treatment and necessary technical improvements for co-producing algal biofuels are described. 1985. capital. the major resource and environmental parameters that Further. focusing primarily on technical and economic performance without adverse the Southwest and southern tier of the United States environmental impacts. and costs.. Lansford et al. and land Sourcing of suitable organic carbon feedstock. Resources and Siting   73 .1). must also be appropriately matched to the required growth Feinberg et al. Assessments of resource requirements and availability and other nutrients—must be appropriately aligned in for large-scale.. water. nutrients. other nutrients. The sustainability and environmental impacts biofuel could be supported if sufficiently high algae of national algae production capacity build-up and productivities could be achieved affordably at scale.. both of which influence siting decisions production efficiency.. to couple land-based microalgae biomass production water. required Photoautotrophic microalgae production energy inputs. the siting and resource factors (e. facilities design. operation over time will be important complementary Many of the findings of these earlier assessments still aspects of the siting and resources issues that will also apply today and the potential remains for biofuels need careful consideration and analysis using tools and other co-products derived from photoautotrophic and methodologies discussed later in chapter 10. concepts are also being proposed. cultivation were conducted during the Aquatic Species and affordability.

whereas photoautotrophic Photoautotrophic Macroalgae Approach production. The latter throughout the country. The Resource issues for the heterotrophic approach are more merits of each should be carefully evaluated.3).2 to organic carbon-rich waste streams. other sugar crops. cost and availability of space 74 9. 2005. Climate CO2 Nutrients Nutrient Recycle Liquid and Gasous Fuels Processing Algal Harvesting and Production Conversion Secondary Products Water Land Climate Water Land Resources and Insolation Location Topograghy Enviromental Temperature Supply/ Demand Use/ Cover Exhibit 9. 1985) can be represented by extension of Exhibit 9. selected to meet production needs. 2006a). A supply issues with first. have different siting and resource input implications and pretreatment of lignocellulosic biomass feedstock and thus present synergistic integration opportunities.g. after sufficient scale-up of production and demand. and land-based ponds.and second-generation biofuels. lead to the Heterotrophic Microalgae Approach problem of linking biofuel production with competing Heterotrophic microalgae biomass and metabolite food and feed markets.. near-shore coastal farms. in terms of cultivation and harvesting. that is expected to be capable of national scale-up to Heterotrophic production can be characterized as more of over one billion tons annually (Perlack et al. beets. and lignocellulosic conceptually include the equivalence of land and biomass. availability. an industrial operation with a significant upstream logistics trail associated with the sourcing of the needed biomass- derived input feedstocks.2 Key siting and resource Parameters Evaporation Salinity Ownership elements for algae biofuel production Severe Storms Allocation Soils Geology (Adapted from Maxwell et al. The algae are adverse food/feed market impacts would be based on cultivated in the dark in closed industrial bioreactors the use of carbon-rich waste streams and the successful that could potentially be established in many locations deconstruction of lignocellulosic materials. Use of sugars from cane. biomass feedstock supply for the downstream value chain. 2005).2 to off-shore macroalgae production.. supply logistics... DOE. Perlack et al. thereby sharing many of the same feedstock culture facilities with off-shore areas and structures. Resources and Siting   . This a photosynthetic origin (Exhibit 9. is more Options for siting macroalgae (also known as seaweed akin to agriculture and serves as the point of origin for the or kelp) biomass production include offshore farms. taking into largely associated with the upstream supply of organic consideration factors such as the scale of farms required carbon feedstock derived from commodity crops. The preferred source of sugars production is based on the use of organic carbon feedstock and other appropriate organic carbon feedstocks for in the form of sugars or other relatively simple organic greatest sustainable scale-up potential and avoidance of compounds instead of photosynthesis. similar conceptual extension holds for the application of Exhibit 9. Achieving affordable scale-up and has the greatest feedstock scale-up potential and is being successful commercial expansion using the heterotrophic pursued and reported elsewhere through bioenergy approach relies on the cost-effective availability of organic programs under DOE and Department of Agriculture carbon feedstock—a resource that ultimately links back to (USDA. Heterotrophic and includes siting and resource issues that are closely photoautotrophic approaches to microalgae production coupled with the production. and from the hydrolysis of starch grain crops can. e.

4 million dry sunlight. the heterotrophic approach can 9. both seasonally and annually. Synergistic Project Intergration Opportunities Ligno-Cellulosic Autotrophic CO2 Sourceess Plant Material algae production . are less sensitive to climate variability than open ponds Based on the scale of macroalgae cultivation practices due to the more controlled environment that closed currently being used to meet non-fuel product demand systems can provide.. research. will most directly metric tonnes for food products).closed PBRs . This is done at the expense of Various climate elements affect photoautotrophic algae 9. the level of production affect productivity. whereas precipitation. evaporation. Heterotrophic algae production A major challenge lies in finding and developing new in closed industrial bioreactor systems utilize mature environments and cultivation approaches that will support bioreactor technology that can provide relatively clean production of macroalgae at the larger scales and lower and protected environmental control needed to optimize costs needed to supply the biofuels CAFO Low-Density Algal . and severe weather will affect water supply and water quality in open systems. Resources and Siting   75 . Closed photobioreactors technologies to facilitate the cultivation of macroalgae. environmental impacts. As illustrated in Exhibit 9. Closed heterotrophic bioreactor Climate systems are decoupled from the variable ambient Sunlight and Temperature Needs outdoor climate. temperature. The co-siting of macroalgal farms with microalgae production. Additional growth conditions for higher biomass density cultures.3 High-level Oil Feedstock illustration of heterotrophic Biomass Residue and photoautotrophic Sugars and Other Heterotropic algae approaches to microalgal Organic Carbon production in Fuels biomass and biofuels Feedstock closed bioreactors production Co-Products and ponds CO2 . and competition production. and day/night light and temperature cycles. has also been proposed as a way of leveraging other evaporation.2 Resources Overview reduce both water and energy-use intensity in the algae production operation. Ligno-Cellulosic N. particularly in the case of land-based with other uses. weather conditions. key other structures such as windfarms (Buck et al. From a resource use perspective.lake. other Water Treatment Deconstruction . Temperature and availability of (annual global production of about 1. and favorable regulatory Heterotrophic algal cultures can be one to two orders of framework for coastal and offshore environment use could magnitude higher in biomass density than possible with help enable cultivation at scales to meet production goals. precipitation.hybrid mix Nutrient-Rich Water. would need to be greatly expanded for biofuels. K.power plants . P.other industrial . rivers Biomass Harvesting or Other Non-Fresh Waters Lignin Dewatering Extraction & CHP Seperation High-Density Algal Biomass FHarvesting Exhibit 9. 2004) factors include solar radiation.4. open photoautotrophic systems that typically achieve no more than about 1 gram per liter culture density.muni WWT & Seperation . and severe weather. technology development.

resource conditions for algae production. and ≥ 200 freeze-free days region 76 9. This could fresh water. i. annual municipal wastewater facilities in the preferred climate average temperature ≥ 55˚ F. but suggest was to provide a preliminary high-level assessment preference for lower latitude. with the major climate element affecting algal It was recognized from the outset that the correlation growth being the availability of sunlight. Wisconsin.e. controls. and the New England states. whereas PNNL study suggests projected productivities in this region could be relatively high.4 presents some simple GIS-based scoping solar insolation. the ocean or sea ≥ 55°F. but the availability of adequate sunlight and the from southeast Texas to northwest Florida is attributed suitability of climate and temperature are key siting and to the lower annual average solar insolation available. identifying preferred regions of the United States for photoautotrophic microalgae production based on Additional factors could conceivably overcome what the application of selected filter criteria on annual might otherwise appear to be uneconomical site and average climate conditions. Resources and Siting   . New Mexico. and marine nutrient might actually be achieved with annual average algae transport cause additional impacts on productivity. from the PNNL study show clear patterns relating climate Texas. Equally important for photoautotrophic microalgae growth however. Arizona. and enclosures requiring geographical regions were: annual average cumulative energy input for environmental control. This is likely due to Preferred Geographic Regions for Algae Production more moderate annual average temperatures despite lower Exhibit 9. Storm and ocean of these annual average climate criteria with what dynamics affecting waves. annual average daily temperature offshore marine macroalgae production. productivity in large-scale deployed systems could only be taken as a gross indicator. the lack of attractiveness of the Gulf Coast region States. For the case of sun hours ≥ 2800. These results should be considered simply conducted by Sandia National Laboratories. The goal indicative at this point rather than predictive.. Within that context. having gross qualitative similarity to Exhibit 9. and the availability of concentrated sources include situations where co-location of inland microalgae of CO2. The more to total biomass growth. The climate criteria used to narrow down the production at higher latitudes might be possible with Exhibit 9.4 Rough scoping assessment of preferred site locations for outdoor algae production a) Regions with annual average climate conditions meeting b) Fossil-fired power plant sources of CO2 within 20 miles of selected criteria: ≥ 2800 hour annual sunshine. Growth of based on the modeling assumptions used. and Florida being most promising). from the standpoint of having adequate solar radiation Projections of annual average algae biomass production (with parts of Hawaii. environment provides a buffer that moderates temperature variation.4 algae is technically feasible in many parts of the United (a). currents. lower elevation sites.4 and the Michigan. would have southern tier states showing greatest productivity potential very low productivity in the winter months. including Minnesota. the availability of non. In Exhibit 9. A significant portion Laboratory (PNNL) using higher GIS resolution and of the United States is suitable for algae production more information-rich climate and meteorological data. California. with the higher growth regions northern latitude states. and annual average freeze-free days ≥ 200. the results are suggestive and consistent with with both open and closed cultivation systems is the the more recent assessment by Pacific Northwest National availability of abundant sunlight. resource factors that will determine economic feasibility.high-cost systems.

Areas with higher annual average and. implications for energy and cost needed to maintain internal temperature sustainable production scale-up. Florida. limited periods of commercially viable algae production. 240 days or more of adequate solar regions of the United States. waves. for closed reactors. and currents that can cause the lower elevation regions of the lower latitude states disruption or damage to physical structures and operations. quality and more costly materials and greater maintenance. throughout the seasons. killing frost in the fall. locations for open pond production. These weather length of growing season are latitude and elevation. For outdoor ponds. The marine environment can also be highly corrosive Texas. the A thorough evaluation of evaporative water loss is conventional crop analog is the greenhouse and the limiting needed to assess water requirements. cooling in the summer. different algal species capable of good productivity in cold season and hot season conditions. For closed photobioreactors. Such analyses should include assessment of the monthly or Water Requirements seasonal solar radiation and ambient temperature ranges. opportunities for utilizing waste heat and energy. The southwestern states (California. will require that suitable climatic conditions loss can be a critical factor to consider when choosing exist for a major part of the year (Maxwell et al. requirements for an open algae growth system. Availability and rotation of Evaporative water loss is discussed in a later subsection. and parts of Louisiana. and hurricanes pose serious concerns in the inland regions of the central states. Evaporation. Areas with relatively long growing prospects for algae production in both inland and coastal seasons (for example. Precipitation affects water availability (both surface it should also establish minimum economically feasible and groundwater) at a given location within a given operational requirement values for heating in the winter watershed region. both high and low. require a more refined biomass productivity and oil content yield. and the Southeast. and New the conventional crop analogy for this is the length of time Mexico) and Hawaii have the highest evaporation rates between the last killing frost in the spring and the first in the United States. Seasonal Considerations are desirable for algae production from the standpoint Various species of microalgae have potential for biofuel of long-term availability and sustainability of water feedstock production and are capable of growing under supply. would Severe Weather Events and Elements provide greater flexibility and could extend otherwise Severe weather events. the Northwest. It is encouraging that researchers today power for cost-effective environmental control of the algae are not only concerned with finding algae with high cultivation. High annual production closely coupled with climate and will increase water for a given species grown photoautotrophically outdoors. Arizona. with more than 60 inches annually. and overall economics. Resources and Siting   77 . Thorough analysis (preferably on a state-by-state basis) with detailed data is needed to assess areas most suitable Water for algae production based on this climate factor. tornadoes. The primary geographical location factors for determining Southwest. discussed later in this section. Evaporation is a Therefore. however. respectively. Offshore marine algae insolation and average daily temperatures above the lower production will be subject to severe weather impacts threshold needed for economically viable growth) are in the form of wind. and coastal areas. hail storms. precipitation (more than 40 inches). dust storms. integrated with co-located industries providing synergistic particularly extremes in temperature. available for productive cultivation. and California. unlike conditions can also potentially lead to increased annual most terrestrial agriculture. 1985). a critical climate issue for both open and closed less important concern for selecting locations of closed photobioreactor systems is the length of economically photobioreactors. Arizona. Other local to materials and usually demands both the use of higher climate and weather conditions will also have influence. such as heavy rain and flooding. although evaporative cooling is often viable growing season(s) for the particular strains of algae considered as a means to reduce culture temperature. of available sunlight per day and the daily and seasonal and need to be taken into account when looking at temperature variations. Discovery and development of algae species capable of increased General Water Balance and Management Needs productivity under wider ranges of light and temperature One of the major benefits of growing algae is that. but also with analysis for systems that would likely be closed and highly algae that grow well under severe climate conditions.industrial operations capable of providing excess heat and locations. of Hawaii. algal culture can potentially average productivities in more geographically diverse utilize non-fresh water sources having few competing 9. Southeast. Evaporative however. Georgia. New Mexico. events can contaminate an open system environment or which have major influence on the hours and intensity cause physical damage to both open and closed systems. represented by specific regions of Hawaii. is a wide range of temperatures.. This would.

Total withdrawals since algal industry scale-up will depend on the geographic the mid-1980s have remained relatively flat at slightly location. In 2005. Hightower et al. depending on the chemistry. Climate change is also recognized as a factor that management issues.uses. Open algal ponds may from the desalination of saline or brackish water sources have to periodically be drained and re-filled. The relatively flat national water withdrawal trend over produced water” from oil. and operational costs. Many of the nations’ fresh ground water aquifers are under up.. along with associated salt build-up and 2008. natural gas. Withdrawals water used for cultivation and the local latitude.S... is used by microalgae (open vs. Some salt byproducts may have commercial net fresh water withdrawals that exceed the sustainable value. 2006b. closed vs.. representing etc. are important areas supply and management in the future (USGS. Kokya et al. or 460. Kenny et al. 2009).000 Mgal/d) and weather conditions (cloud cover. elevation. Hutchison et use. This will require periodic blow. 2009).S. reflects for open pond systems in more arid environments with the fact that fresh water sources in the Untied States are high rates of evaporation. 2002). 2009). can also become an environmental problem for inland where a growing number of counties are experiencing locations. the application of desalination treatment to control increasing stress. withdrawals.. total combined production operations could be for the replacement of water fresh and saline water withdrawals more than doubled continuously lost to evaporation from open cultivation from about 180 billion gallons per day in 1950 to over 400 systems. USGS. Whether or not this is a problem for sustainable billion gallons per day in 1980. nearly and the site-specific details of climate. ). such as saline and brackish ground water. water demand over the 30 years prior to that. Kenny et al. This will be of greatest impact and concern in being fresh (Hutson et al. At the national scale. Mao. humidity. for future research. the thermoelectric-power industry in the coastal states whether evaporative cooling is used for closed systems. 2006b. Irrigation withdrawals represented 31% of all water al.000 heterotrophic). Saline water (seawater and heterotrophic assessment. thereby changing fresh water supplies in support of development and the culture conditions. Oroud. Growing demand for limited will change with evaporative water loss. from both a resource perspective could have major effect on all sectors of water resources and an algal cultivation perspective. various locations throughout the eastern half of the country. Implementing water desalination would impose additional The stress on fresh water supplies in the United States is capital. humidity. with water use in upstream organic million gallons per day (Mgal/d). water-sparse locations. and 95% loss in open systems will be the degree of salinity of the of all saline-water withdrawals in 2005. and the future expansion of fresh water salinity build-up. or “co..). about 95%. However. periodic addition conservation and by the increased re-use of wastewater. 2002. Water use will also depend on brackish coastal water) withdrawals were about 15% of the type of growth systems used for photoautotrophic the total. and coal-bed the past 25 years. 1999. 2009). 2008. to cool electricity-generating equipment. different dominant algae species and growth conditions. Mgal/d and were the second-largest category of water and wind conditions (Al-Shammiri. 1978. following the more than doubling of methane wells (Reynolds. or highly adaptive algae that can thrive supplies for non-agricultural use must increasingly come under widely varying conditions. and locally available water over 400 billion gallons per day.. 2003. solar insolation. Resources and Siting   .000 acre-feet carbon feedstock production needing to be part of the per year (Kenny et al. one-half of the total U. population increase has thus far been offset by increased down of ponds after salinity build-up. climate conditions.. were for thermoelectric-power generation. Scoping Out Water Requirements for Algae Production Analysis of U. 2004. from pond but is also becoming a local and regional concern at drainage and replacement. Water Supply and Management Water use and consumption for algae-based biofuels will Total combined fresh and saline water withdrawals in the depend on the cultivation approach (photoautotrophic/ United States as of the year 2005 were estimated at 410. 1995). 61% of all surface water. management. 2009). or staged as a and from the treatment and reuse of wastewater. of non-saline make-up water to dilute the salinity build. modeling. energy. 2008. or from desalination operations. Also a complicating factor for evaporative water 41% of all freshwater. and field assessment. Almost all saline water. all of cascading sequence of increasingly saline ponds each with which have increasing energy demand implications (DOE. withdrawals (201. for irrigation of crops and other lands totaled 128. salinity and water chemistry approaching full allocation. wind. solar insolation. hybrid combination). and 37% of all freshwater withdrawals A significant source of water demand with inland algae (Kenny et al. which also tend to be in the more arid and higher solar resource regions like the Southwest. Disposal of high not restricted to the more arid western half of the country. salt content effluent or solid byproducts. A rough upper bound estimate of evaporative water loss 78 9.. Hightower et al. Water balance and supply from precipitation (DOE.000 ambient temperature variations. with the majority (85%) resources.

Saline groundwater resources. Along with geological data. by state engineers and water authorities. and to predict can be significantly reduced if closed systems are used. in gallons of water per gallon of algal biofuel produced. discussed be an over-estimate of what can be expected under later in this chapter. with open systems. Additional analysis.7). southwestern United States. integrating algae pan evaporation data will be worst-case and will likely production with wastewater treatment. 1982a and 1982b. and residential development needs in high growth rate Water must be considered a key element of life cycle water-sparse regions like the Southwest (Clark. depending on systems and process specifics. reclaimed. There is considerable in the United States are areas of investigation in need untapped potential for utilizing brackish. Dennehy. additional water will be used and increasing interest and potential competition for access as consumed. limited fresh water supplies. the estimated capacity. Resources and Siting   79 . 2007. and may well also be saved. Developing and Mapping Water Resources for Algae Production The evaporative loss estimates from the notional scenarios When considering the water resources needed for the provide an indication of the potential magnitude of the future development and expansion of algal biofuel issue. putting less additional demand on evaporative loss than fresh water. analysis for algal biofuels.. where brackish or saline groundwater supplies may be The most optimistic production scenario was for the both ample and unused or underutilized. The notional scale-up scenarios and assumptions made are discussed later in this chapter (see Exhibit 9. 2009). which assumed annual quantitative information remains limited on U. Extrapolations based on fresh water From a resource use standpoint. Hightower et al. Shuttleworth.. depth information determines the cost of drilling and operating In summary. and sustainable withdrawal optimistic productivities are assumed. and sustainable withdrawal rates Evaporative water loss associated with algae cultivation for these non-fresh ground water resources. water utilization for algal biomass and (including energy input requirements for pumping) a downstream production of biofuels. the use of non-fresh water sources will need open horizontal pan evaporation data (Farnsworth et to be emphasized in the face of the growing competition al. 1984) to large area scale-up of algae cultivation (DOE.S. systems are implemented. define the spatial distribution. depth. 2007. and of greater investment. resource development. has the potential benefits of actual operating conditions in the field. An improved knowledge base is needed to better evaporative water loss intensities will be greater.. 31 g/m2 per day with 50% dry weight oil content. as noted earlier. the effects of their extraction on the environment (Alley.with open systems was done as part of a hypothetical co-produced water. both in terms of well in a given location (Maxwell et al. and evaluated under a depth. other inland could be a consequence of open system operation parts of the country can be targeted for algae production in the more arid and sunny regions of the country. or several hundreds of non-fresh water over a range of salinities means that. 2009). and were based on simply applying fresh water production. physical and chemical characteristics. Water requirements of water for algae production than deeper sources located information will not be well characterized until larger scale longer distances from the cultivation site. 2006b. saline. Woolhiser and demands on limited sustainable fresh water supplies et al.. and chemical characterization of saline aquifers range of site locations and conditions. 2008). 2004). The maps of saline groundwater 9. warrants aquifers located closer to the surface and nearer to the closer attention and assessment to better understand and cultivation site would provide a more cost-effective source refine water resource requirements. monitored. Evaluation of water use for the overall value chain from 2003. If less water quality and chemistry. are also of Along the way. Suitable overall input supply needs and consumption. brackish average algal biomass and oil productivities of nearly and saline groundwater resources in terms of their extent. 1993. 1985). Open bodies of productively using non-fresh wastewater resources brackish and saline water will usually experience less for renewable fuels. Gerbens-Leenes et al. and recycling nutrients. The evaporation estimates suggest that water loss on the order of several tens of gallons of water per kilogram The unique ability of many species of algae to grow in of dry weight biomass produced. addition to coastal and possible off-shore areas. NAS. Unfortunately. and a source of water for treatment and use to meet commercial recycled. experimental scale-up scenario study in preparation for the National investigation. quantity. needed to understand how best to leverage these resources. The location. and field trials at larger scales of operation are Algae Roadmap Workshop. algal cultivation through harvesting and post-processing particularly deeper aquifers that are largely unregulated into fuels and other products will also be important. as with other forms of Depth to groundwater is pertinent to the economics of bioenergy (NAS. reducing eutrophication of natural water bodies. held in December 2008..

In represented by the algae biofuels and other co-products. 2009. description of substances toxic to algal growth) and the local availability and distance to suitable land for algae 80 9. at some large growth system and incident light conditions. Flue gas. NATCARB. Resources and Siting   . recharge rate. transport. Larger coal-fired base-load generators that locate. Dennehy. However. Campbell carbon capture and reuse in the form of fuels and other et al. sustainability of supply.. Applications fuel-fired power plants. detailed analysis is difficult estimated 20% to 30% of the total power plant emissions and often lacking. USGS. represent over half of the more than 6 billion metric tons rates and quantities of CO2 produced. the utilization of fossil carbon emissions and complement are already in operation today and under consideration for subsurface sequestration. algae production does possible broader use for carbon capture and storage (CCS) not actually sequester fossil carbon. 2005. separating CO2 in large industrial plants. typically output a steady 1. identify. content. 2008b). gas abatement credits would come from the substitution The rate of effective CO2 uptake will also vary with of renewable fuels and other co-products that displace or the algae species. but rather provides in response to climate change (Rubin. Likely Stationary CO2 Emission Sources Major stationary CO2 emission sources that could An inventory of stationary industrial CO2 sources in potentially be used for algae production are shown in the more promising regions of the country. information is needed to improve our understanding of this resource in support of algae production siting Barriers to Viable CO2 Capture and Utilization analyses (Dennehy. Power generation alone (mainly developed by the USGS prior to the mid-1960s (Feth. including natural Algae production could provide excellent opportunities for gas treatment plants and ammonia production facilities.5. including algae production. The CO2 generated by the power (Shannon. As a result. A limited plant can only be effectively used by the algae during amount of this information is available for major the photosynthetically active sunlight hours.. and of CO2 emitted annually in the United States (EPA. and coal bed methane be captured and used affordably for algae production wells is another closely related and underutilized water will be limited by the operational logistics and resource that can range in quality from nearly fresh efficiencies. or more than 1965. and details of reduce fossil fuel consumption. Photoautotrophic algae will only utilize products derived from the algae biomass.000 to 2500 MWe of power would each require many tens of thousands of acres of Carbon Dioxide algae production and large volumes of water to provide a similar effective offset of 20% to 30% of the CO2 emitted. More detailed and up-to-date 2 billion metric tons per year (EIA. and analyze this information. potential As an example.. 2002). 2004). Produced The degree to which stationary CO2 emissions can water from petroleum.. Therefore. for algae cultivation scale-up within reasonable geographic proximity of stationary sources. and quality production to fully utilize the CO2 in the flue gas emitted (chemical components and characteristics) are important from a 50-MWe semi-base load natural-gas-fired power information for assessing non-fresh groundwater aquifer plant would require about 2. natural gas.g. it will be necessary to provide a CO2 source that is suitably free of materials potentially toxic to algae. using coal) represents over 40% of the total. but a significant effort will still be needed to et al. In addition. 2009). The geographical location.resources are based on incomplete data that was largely EIA. The Carbon Capture Opportunity in Algae Production Efficient algae production requires enriched sources of CO2 The distance for pumping flue gas to algae cultivation since the rate of supply from the atmosphere is limited by systems will become a limiting factor that requires diffusion rates through the surface resistance of the water capture and concentration of CO2 from the flue gas for in the cultivation system. 2006). such as from fossil. is not a simple issue. biomass growth rate. addition. scale of algae production. local aquifers may be due to CO2 off-gassing during non-sunlight hours and the available through state agencies and private engineering unavoidable parasitic losses of algae production (Brune companies. aquifers. and distribution could require more production. longer distance transport and distribution. parasitic losses from flue gas the requirements for CO2 supply to enhance algae treatment. 2008 and 2009). and the matching of CO2 source availability energy input than the output energy displacement value with algal cultivation facilities. 2008). if these aquifers are spread over the greenhouse gas emissions offset will be limited to an large geographic areas. collect. 2008 and 2009). 2008) characterization of the CO2 emissions stream (e.200 acres of algae cultivation resource availability and suitability for economic uses area (Brune et al. depth. spatial extent. a recent analysis suggests that for algae yield. and the availability of land and water to hyper-saline (Reynolds 2003. The sources shown (NATCARB. would be a good source of CO2. 2004. However. 2009). Data on small. Any greenhouse CO2 during daylight hours when photosynthesis is active. including Exhibit 9.

is needed for making refined assessments for Land availability is influenced by various physical. grassland. Soils. One outcome economic.3 112 Electricity Generation 2. several characteristics need to be considered. as illustrated in of a hypothetical algae production scale-up scenario is Exhibit 9.0 13 Industrial 141.1 4. R&D program (e.g.5) within practical transport distances of suitable algae and relatively barren desert land. much of the land in the West is government owned. 2006 and 2009). social.2 475 Gas Processing Refineries/Chemical 196.6. lease. and political factors. as is expected requires relatively flat terrain.production. Exhibit (USDA.g.. and particularly their porosity and permeability Land characteristics. 9. Publicly and privately owned lands are algae culture surface area to produce 10 million gallons of subject to variable use.9 173 Total 3. including pasture. For a realistic appraisal production sites in a given geographical region. This of land for algae production (i.002 Ethanol Plants 41.e.276. affect the construction costs and design Factors for Evaluating Land for Algal Production of open systems by virtue of the need for pond lining or Land availability will be important for algae production sealing. oil feedstock. Physical characteristics.3 163 Fertilizer 7. Areas with more than 5% with any photosynthesis-based biomass feedstock. legal.9 665 Other 3. Resources and Siting   81 . Topography would be a limiting factor for these because either open or closed systems will require systems because the installation of large shallow ponds relatively large areas for implementation. Hundreds of millions of acres of relatively low- the limited quantity of CO2 that would likely be available productivity. such as topography and soil. lower-value land exists in the United States from stationary industrial point sources (e. which means that environmental assessments and/ Exhibit 9. For example. any given region unless other factors drive the investment in expanding the nation’s CO2 pipeline infrastructure. algae production siting and CO2 sourcing. land that would be both can be expected to constrain the extent to which algal suitable and potentially available for siting algae production biofuels production can be affordably scaled up within facilities). 2008a) CO2 EMISSIONS CATEGORY NUMBER OF SOURCES (Million Metric Ton/Year) Ag Processing 6. and purchase requirements.5 3. target annual average biomass production of 30 to 60 g/m2 per day with 30% to 50% Land ownership information provides valuable insights neutral lipid content on a dry weight basis). could limit the land available for open pond algae farming.3 140 Cement Plants 86.5 Major stationary CO2 sources in the United States (NATCARB.796 9.6 53 Petroleum and Natural 90... Even slope could well be eliminated from consideration due to at levels of photoautotrophic microalgae biomass and oil the high cost that would be required for site productivity that would stretch the limits of an aggressive preparation and leveling. such systems on which policies and stakeholders could affect project would require in the range of roughly 800 to 2600 acres of development.702. as will be discussed further in chapter 10.

or environmental impact statements could be required restrict the fraction of incident photon flux that can be as part of the approval process. 2009). It must be stressed that “potential” will be constrained by the available energy density algal biomass and bio-oil production projections at in sunlight and the relatively low efficiencies of commercial scale currently remain hypothetical rather photosynthetic processes coupled with other systems losses. Resources and Siting   . given that large-scale algae biomass production The result will be theoretical and practical upper limits on intended for bioenergy feedstock does not yet exist. the cost of land is during hotter ambient conditions. than real.. Loss mechanisms that often not considered or is relatively small compared to other capital cost. This the light and shade their neighbors farther from the light is significantly lower than the more optimistic target source.60 g/m2 per day. as with any form of biomass. (chapter 10). the potential remains for high algae biomass production relative to more conventional Further. lower-value production with algal strains otherwise be effectively captured and used by the algae developed and improved over time to be more suitable and in photosynthesis are instead converted to thermal (heat) productive for biofuel feedstock rather than those used energy in the culture media and surrounding cultivation for today’s high-value algae biomass product markets.. the amount of biomass growth that can be achieved per unit Relatively large-scale commercial algae production of illuminated surface (Weyer et al. such systems have not been optimized for energy losses that occur when incident photons that cannot higher-volume.20 g/m2 per day. 1985 ) 82 9. the dissipative heat loading Land Availability Constraints can be a benefit in moderating culture temperatures and Land use and land value affect land affordability. reviewing the more recent economic analyses for algae or can lead to overheating and loss of productivity biomass and projected oil production. Indian reservations also effectively used to drive photosynthesis ultimately places comprise a significant portion of this land.6 Factors and process steps for evaluating and constraining the available and appropriately suitable land for algae production (Adapted from Maxwell et al. Land ownership practical upper limits on the biomass productivity.. for algae productivity. 1985). algae productivity crops. system structures. Depending on the algae strain and culture system approach used. By improving productivity under colder ambient conditions. Optimizing light utilization in algae production projections for biofuel feedstock of 30 . Contributing to with open ponds for high-value products can serve as a productivity limits per unit of illuminated surface area is the baseline reference. but currently reflect lower biomass fact that algal cells nearest the illuminated surface absorb productivities in the range of 10 . Land that is highly desirable for development and other set-asides for publically beneficial reasons may Typography Land Ownership Current Land Value Systems Engineering All Physically Politically Land Suitable Affordable Available Available Parcels Land Land Land Land Enviromental Restrictions Current Land Use Legislation Water and and Regualtions Climate Resources Exhibit 9. systems includes the challenge of managing dissipative However. can represent and impose political and regulatory Despite the practical upper limits that will naturally exist constraints on land availability (Maxwell et al.

wastewater treatment and recycling • Potential to be integrated with power plant must be incorporated with algae biofuel production. excess nutrients (e. archaeological sites. or other CO2-emitting industry operations The main connections of algae production and • Potential to treat agricultural drainage wastewater treatment are the following: and eutrophic water bodies • Treatment technology is needed to recycle nutrients and water from algae biofuel processing residuals for use in algae production. biomass production facilities 1985). The imported wastewater Wastewater Treatment and would be treated as part of the algae production.g. and potassium) be a key issue for achieving affordable and Algae can be useful in the treatment of waters polluted with sustainable scale-up of algae biofuels production. Examples of this type of constraint include technologies. and potassium). and wastewater treatment facilities. The roles of nutrients. biofuels industry are discussed in this subsection... Resources and Siting   83 . phosphorus. On the other hand. and the algae production can algae feedstock production facilities. as noted earlier. which are and processing could be made up with wastewater used to dispose of wastewater or brines. help provide nutrient removal service in the wastewater For large-scale algae biofuel production. particularly in the eastern half of the deployed in the near-term and provides workforce country. which can also help supplement and off-set these facility types in the development of an algae the cost of commercial fertilizers for algae production. offsets algae production costs • Lower capital and O&M costs than 9. Beyond economics. Land cover categories such as barren and scrubland cover a large portion of the West and may • Development of skilled algae production workforce provide an area free from other food-based agriculture • Potential for nutrient recycling at algae where algae growth systems could be sited (Maxwell et al. The same applies to land highly suited for higher-value agricultural POTENTIAL BENEFITS OF ALGAE PRODUCTION use. The availability and sustainability of water supplies • Wastewater treatment revenue that in the West will also be a key consideration. • Imported wastewater provides nutrients and water to make-up inevitable losses. nutrients from treatment. Nutrient which produce algal biomass as a consequence of the recycling would be needed since wastewater flows in wastewater treatment. Sensitive environmental or cultural land constraints Although algae-based wastewater treatment requires will also reduce the overall land availability (Maxwell many times more land area than mechanical treatment et al. Two main types of algae production facility are wastewater (municipal and agricultural) would be captured envisioned: dedicated facilities. dairy and feedlot operations located throughout the • Algae-based wastewater treatment can be United States. phosphorus. 1985). for additional rounds of algae production. some land • Early opportunity to develop large-scale cover characteristics could present excellent opportunities algae production infrastructure for algae farming. organic matter. nutrient recycling. Recycling Applications • Algae-based wastewater treatment provides a needed Municipal wastewater treatment facilities and agricultural service.4 Integration with Water conventional wastewater treatment Treatment Facilities • Lower energy intensity than conventional wastewater treatment (a greenhouse gas benefit) Inevitably. Dedicated biomass production the United States are insufficient to support large-scale facilities will also require wastewater treatment and algae production on the basis of a single use of nutrients. wildlife areas. represent potential co-location sites for algae training and experience in large-scale algae operations where nutrient-rich wastewater could be cultivation that would translate to future dedicated used for algae production. with the main purpose of by algae and then recycled from the oil extraction residuals biomass production. algae-based parks. treatment has the following advantages: and other historical sites. monuments. A subset of wastewater treatment Inevitable nutrients losses during algae production facilities consist of evaporation facilities. metals. in suitable climates.. synthetic organic compounds. Supply and cost of nutrients (nitrogen. nitrogen. 9. this also avoids the perception WITH WASTEWATER TREATMENT and potential conflict of food and feed production versus fuel.not be seen as suitable for algae production.

Thus. harvesting is a 2002).. The bioaccumulation of trace contaminants in high-value products (>$100/kg biomass).7. but research is needed to improve the reliability of these processes (as Algae-based treatment facilities are typically less expensive discussed in chapter 4).. which is algae that would occur in the receiving waters. dissolved oxygen. the major types to build and to operate than conventional mechanical of wastewaters available for combined algae production treatment facilities. food processing).. Mechanical treatment technologies typically hold either high rate ponds (~30 cm) or algal turf scrubbers the wastewater for less than 12 hours. Closed photobioreactors are not emphasized technologies hold the wastewater for at least several days in this wastewater treatment discussion since they are and in an environment similar to many natural receiving likely to be economical only when also producing waters. followed by algae separation in dissolved treatment are aligned. Finally. autoflocculation) are far less costly. to oxidize waste organic matter (Exhibit 9. Even with internal recycling.. The major plants in that they both produce oxygen and assimilate classes of wastewaters to be treated are municipal. high-productivity algae and water treatment are those contaminated with organic ponds have a total cost that is about 70% less than activated matter and nutrients (e. As noted above. Algae are similar to in conjunction with wastewater treatment. agricultural drainage. 1988.g. These highly productive systems use shallow reactors. organic nutrients. The cost biomass production.g. As with other algae production systems. These reactions are also the best-known industrial (e. whereas pond (~1 cm). The great depths contribute to low algae organic compounds such as phenol and acetonitrile productivity. Despite a seeming of algae to assimilate dissolved nutrients down to trace abundance of wastewater and waste nutrients. 2002). Muñoz et al. recycling of concentrations is useful in wastewater treatment. municipal and industrial sources) sludge. Resources and Siting   . 2002. organic agricultural mechanisms of wastewater treatment by algae.. The standard of nutrient removal and wastewater treatment. might be prevented to a 84 9.17 per m3 treated) or manipulation of culture conditions to promote lipid (Maglion.. Non- production. For example. 1998. 2008) is high for biofuel production.and potentially endocrine disrupting compounds (Oswald. The ability agricultural drainage. Heavy metals may be removed Algae Production Techniques for by adsorption to algal cells.7).... including nutrient removal. Maintenance of lipid-rich strains. This nutrients (e. practical opportunity to install algae production facilities requiring two types of treatment. lakes. Existing interaction of algae and bacteria in wastewater cultures algae-based treatment facilities use relatively deep leads to degradation of a wide variety of synthetic ponds (1-6 m).g. eventually unlikely when wastewater contaminants are present.10 . at least in terms of maximizing air flotation units or sedimentation clarifiers. 2009).$0. Ponds for more advanced compounds such as human hormones and antibiotics treatment. 2009. including wastewater treatment ponds. need high algae from animal facilities) is an area in need of study. natural disinfection and remove trace contaminants. confined animal facilities).g. which is the leading water treatment technology and wastewaters mainly contaminated with inorganic used in the United States (Downing et al. The (e. provides a Organic-rich wastewaters usually also contain nutrients. Aksu.. and naturally occurring organic catalysts (Sinton et al. and eutrophic waters dissolved oxygen algae release is used by treatment bacteria with low organic content but high nutrient content (e. 2008) and throughout the world. rivers.. endocrine disrupting matter and pathogens only). harming higher organisms.g. and rivers). needed if algae are to make a substantial contribution to Less well-known are the ability of algal systems to provide national biofuel production. The treatment goals of these facilities (removal of organic removal of trace contaminants (e. the method is chemical addition to achieve coagulation and objectives of biofuel feedstock production and wastewater flocculation. of chemical addition ($0. which will be a benefit as Wastewater Treatment Plants long as the resulting metals concentrations in the algae biomass are not excessive or inhibitive for later use in Integration of algae production with wastewater treatment the processing of fuel and other co-products.. and lakes). to make up for nutrient losses (Brune et al. Kohn et al. if the nutrients and carbon at algae production facilities will be nutrient-rich algae are then also removed from the water. cost savings. 2007). importation of wastes and/or wastewater will still be Disinfection is promoted via the production of oxygen needed in dedicated algae biomass production facilities radicals in the presence of sunlight.. 2003. 2005).g. High rates of algae production lead to high rates crucial step in wastewater treatment systems. coupled with the tremendous need for expanded and improved wastewater treatment in the United Treatment of Organic Wastewaters for Algae Production States (EPA. but high productivity is not crucial to the (Borde et al. Borde et al. Woertz et al. productivities (as does biofuels feedstock production).. the is illustrated schematically in Exhibit 9. have yet to be demonstrated consistently chemical flocculation processes (bioflocculation and for ponds.

Resources and Siting   85 .Exhibit 9. High Rate Clarifier Clarifier Pond Raw Sewage Treated Effluent Sludge CO2 Digestor Generator kWh Food Algae Biomass Production Inputs Option Liquid Biofuel Products Reclaimed Water Fuels Fertilizer To Fertilizer b) Simplified conceptual system block diagram 9.7 Integration of algae production with wastewater treatment for nutrient removal and biomass production: Sun CO2 Reclaimed Algae Water CO2 N P Waste CO2 O2 N Biomass Water P Organics Bacteria a) Basic principles of operation.

2004)... (GHG) emissions avoidance benefits compared to conventional wastewater treatment technologies. Furthermore. Thus.. Treatment of nutrient-rich National Algal Roadmap Workshop break-out sessions.5 Co-location of Algal nutrient. micro-screening. 9. 2002). low-organic content wastewaters will expand Cultivation Facilities with the opportunities for algae production systems. an overriding or other wastewater. However. CO2-Emitting Industries Agricultural drainage and eutrophic water bodies (e. The processing of the algal biomass for fuel • Develop algae grazer control strategies and other co-products would presumably destroy and • Develop reliable low-cost algae harvesting neutralize the contaminants.. constraints significant. Mulbry et al.) are examples of such waters This section includes findings from discussions held at the (Benemann et al. Findings from rate ponds with CO2 addition has been demonstrated in these interviews and conference calls were integrated California’s Central Valley and elsewhere (Craggs et with the Workshop inputs in developing this section. It is important to point out that amongst the numerous barriers to co-location of algal cultivation facilities with High rate ponds might be used as components of industrial CO2 sources identified at the Workshop and evaporation systems needed to dispose of blow-down subsequent discussions with electric utilities. 1996. but further investigation is techniques. hazards to wildlife from toxic compounds (e. such as bioflocculation. market from decomposition of organic matter would not be drivers. technical and market challenges. For all biofuel wastes for algae production feedstocks. CO2 addition or slow atmospheric of discussion included the value proposition. mine drainage. Treatment of agricultural drainage with on large-scale development. potentially increasing • Demonstrate recycling of biofuel processing the cost of waste disposal or recycling. regulatory Successful use of high rate ponds specifically for and economic) for algae facilities that are co-located nutrient removal/recycling requires resolution with electric utilities and other industrial CO2 sources. al. needed to confirm this.g. Treatment of Inorganic Wastewaters for Algae Production In addition to the ability of algae systems to treat organic-rich wastewaters. and included For algae-based treatment of low-organic content several large municipal electric utilities.. Calif. desired absorption is essential since inorganic carbon generation outcomes. of several issues. routes of such contamination should be • Determine the cost savings and greenhouse gas studied and preventative measures developed. Salton Sea. The high rate ponds could create theme was that electric utilities primarily view algae an algal product while performing the service of water cultivation as a means of CO2 capture as opposed to a evaporation. potentially leading electric utilities through later meetings and conference to greater land availability and savings in land costs. research efforts and policy evaluations would likely need to focus on Main Research Needs for Algae both carbon capture and the production of biofuels and Production with Wastewater co-products to overcome barriers (technical. waters is likely to occur in more rural settings than and additional input sought from managers at major treatment of municipal wastewaters. contaminating the algal biomass likely would remain in the waste from biofuel processing. Evaporation ponds are currently used to method for producing biofuels and co-products.. and potential algal turf scrubbers without CO2-addition and high opportunities for the federal government. dispose of agricultural drainage. integration opportunities and challenges. very different business models and goals for algae selenium.g. The topics wastewaters. as follows: • Large-scale (3-5 acre) demonstration of CO2- enhanced high rate ponds for nutrient removal • Determine CO2 biofixation efficiency 86 9.great extend by pond treatment followed by algae • Determine growth model parameters harvesting. co-products. oil field produced electric utilities may need to partner with algae cultivation/ water. etc. any heavy metals autoflocculation. These follow-on efforts were coordinated with the Electric Power Research Institute (EPRI). 2008. chromium) must be carefully evaluated. their ability to treat high. etc. As with any evaporation pond technology companies and fuel refiners/distributers with system. production in order for this type of co-location to be widely commercialized. Lundquist et al. Resources and Siting   . calls.

This higher concentration would affect Coal-fired power plants may be a convenient source of the sizing and operations of algae production facilities— CO2 for algae production. Resulting costs for • Excess heat or power may be available to provide CO2 will be site-specific and dependent on methods of heating or cooling for improved thermal capture.5. depending on the type of plant and fuel used. construction of algae systems at natural models described in more detail in the Systems and gas-fired power plants may be a better investment. The impact on algae production would depend quantitative breakdown. The emissions from fuel in lieu of permanent sequestration. but from an emissions control an aspect that could be incorporated into engineering perspective. introduced earlier in Exhibit on the phasing of the intermittency with respect to the 9.). beneficial re-use of cooling water and wastewater. and distance of transport to algae management of algae cultivation systems – this cultivation sites. Resources and Siting   87 . the CO2 would be emitted to the atmosphere if not captured and sequestered by other means (Rubin. The growth. The Techno-Economic Assessment section of this report. coal-fired plant flue gas typically expected to result in a net reduction of overall GHG has about a factor of two greater CO2 concentration emissions. plants – overcoming a primary resource challenge for algae cultivation at scale. and delivery of CO2 from the power plant to of this will need to be addressed before utilities the algae cultivation site. which typically ranges from about 5% to about 15%. However. carbon sequestration. concentrated • Abundant quantities of concentrated CO2 available sources flue gas from fossil-fuel burning power plants from stationary industrial sources can supplement and other CO2-emitting industrial sources can be low concentration CO2 from the atmosphere.. which can transportation fuels may not rigorously be considered bring some advantage in terms of efficiency of capture. fired power plants. beneficially used in algae production. but economic viability under a broader range of geographic and climate must be determined case by case.6%). generators will have intermittent operation that would A variety of stationary industrial sources of CO2 introduce intermittency in the supply of CO2 for algae are distributed throughout the United States. as sources.The Opportunity in Co-Locating with CO2-Emitting Sources ADVANTAGES OF CO-LOCATION OF Since photoautotropic algae growth requires CO2. it is also relevant to other CO2-intensive • Potential carbon credit for utilities. A number of large coal-burning power plants with baseload coal-fired plants. shows that fossil-fired power plants represent daylight hours when photosynthesis is active. many of these facilities have higher CO2 concentrations compared to power plant flue gas. The regulatory implications transport. Co-location of algal cultivation with industrial available. 9. cement manufacturing. fermentation-based industries. Gas-fired the majority of CO2 emissions from stationary baseload generators would not be intermittent. while providing While the information in this section focuses on fossil. in that the carbon will be require proportionally larger algae production systems re-released to the atmosphere when algal-derived per unit energy produced and higher costs per unit energy fuels are combusted. would also emit CO2 distributed across the southern tier states provide during periods of darkness when it cannot be utilized by ample sources for algal growth on a large scale. gas-fired power plants and fuel companies are likely to widely adopt algal that operate as peaking plants rather than base-load cultivation co-located with industrial CO2 sources. fossil fuel require establishing a U. but.15%) than natural gas plants (5 . Offset of fossil fuel conditions on or near a year-round basis. policy on carbon extraction/refining. found often in proximity of power CO2 sources is a promising area for further research. some absorption and re-use as transportation geothermal power production. the process of capturing flue-gas CO2 to make (10 .S. etc. 2005). Also. and ALGAE PRODUCTION WITH STATIONARY productivity can be enhanced by supplementing the INDUSTRIAL CO2 SOURCES limited CO2 available from the atmosphere. using algae to capture the maximum of CO2 absorption by algae in any carbon-credit or amount of CO2 emissions from coal-fired plants would cap and trade framework. This will industries (e. While algae biofuels can be produced. During those times.g. reason is that coal-fired power plants have higher CO2 emissions per unit energy produced than natural gas-fired An important policy question to consider is the value power plants. consumption by algae biofuel and other co-products must be done with approaches that provide a net GHG emissions • Excess wastewater or cooling water may be reduction. Thus. conditioning. the algae through photosynthesis. Costs are expected to be lower than for will allow developing algal cultivation facilities pure commercially supplied CO2.

the uncertainty may pose a barrier for wide commercial Directions for Research and Development adoption of the technology. solar resource. or in some cases algal to capture CO2 as opposed to producing biofuels and cultivation could be co-located with both stationary co-products. The use of flue gas to cultivate algae will involve non-point source emissions at ground level. as well as policy-development • Land availability: Suitable and affordable vacant land efforts. Thus. These costs and losses must be parasitic losses of CO2 transport. Although often referred to as a “free” resource. Specific test-bed R&D absorbed during periods when sunlight is available topics relevant to power and wastewater utilities and photosynthesis is active in the algae. ethanol plants. the capture and delivery of concentrated CO2 from The test-beds would ideally be located at power plants. minimized and compared to other approaches for the ▪▪ Evaluation of temperature control (power capture and sequestration or reuse of carbon. and be absorbed by algal ponds at a realistic scale climate conditions. distributors. and facilities that will aid the following: are based upon point-source emissions from high ▪▪ Rapid and consistent engineering design elevations. This effort could involve a likely to be possible near the power plant consortium of R&D organizations. The same generally holds true for other cultivation companies. and new business models will be needed to • Unclear regulatory framework for carbon-capture commercialize this approach. algal technology companies.Barriers to Co-Location of Algae • Maintaining cultivation facilities during utility outages Production with Stationary Industrial and through seasonal changes in algal growth rates: Detailed models will be needed to develop and evaluate CO2 Sources approaches for managing the variable nature of both • Need for nutrient sources: While stationary industrial CO2 emissions and algal growth rates/CO2 uptake. Therefore nutrients must be production arena.10 acres). to conduct research at the pilot scale (3 . advantages to capture and deliver industrial CO2 to ponds and of decentralized production considering grow/harvest algae. stationary industrial sources of CO2 (cement refiners.30% of a power plant’s greenhouse gas emissions can be offset ▪▪ Development of efficient test-bed facilities by algae biofuel and protein production (Brune et al.. Also. etc. will be required for commercialization of may not be available adjacent to or near major algal cultivation facilities co-located with industrial power plants CO2 sources and/or wastewater treatment facilities. and other participants coordinated plants. algal facility. credits: Until there are regulations in place that quantify carbon credits from algal growth facilities. ethanol plants or other and optimize algae production will not be “free”. sources of concentrated CO2 can potentially provide • Resistance from electric utilities: Electric utilities are ample carbon for photosynthesis-driven algal growth. Several areas for research. required. universities. not in the fuels business and regulated public utility in most cases there will not be a complementary commissions will be constrained in entering the fuel nutrient (N. etc. CO2 is only at the national level. mechanisms to encourage partnering CO2 sources and nutrient sources such as wastewater between utilities and algae/fuel companies will be treatment facilities and agricultural waste streams. Resources and Siting   . K) supply. ▪▪ Techno-economic analyses • Capital costs and operational costs: There exists a ▪▪ Life Cycle Analysis and GHG abatement analysis need to evaluate capital costs and parasitic operational ▪▪ National inventory of potential production sites losses (and costs) for infrastructure and power required ▪▪ Evaluation of economies of scale vs. include: ▪▪ Technology evaluation at larger scales ▪▪ Determination of algae production facility model parameters 88 9. • Emissions from ponds are at ground level: Regulatory The following directions have been identified: requirements from power plants and other stationary • Develop computer models of algae production sources are governed by the Clean Air Act. Their fundamental objective will be brought in from other sources.). P. and agricultural • Too much CO2 near plants for realistic absorption: drainage/water body restoration sites that also represent Large power plants release too much CO2 to a range geographical locations. Current plant cooling and algae pond heating) estimates are that approximately 20% . stationary industrial sources as a supplement to enhance wastewater treatment facilities. • Establish national algae biomass production test-beds 2009). CO2 emitting industry facilities.

flocculation harvesting. etc. and refiners/distributors. etc. costs. between public utilities/other industrial CO2 sources ▪▪ Algal biomass handling. proteins: carbohydrates and C:N:P) ▪▪ Power plant cooling with treated wastewater in ▪▪ Methods of nutrient and water recycling within conjunction with algae production production facilities. storage. pathogen safety • Develop and train the future algae production/algae ▪▪ Beneficial management of residuals for soil carbon biomass processing workforce at the national test-bed development. ▪▪ Development of algal strains and their cultivation techniques 9. salinity and blowdown • Evaluate policies that would encourage partnering management. Resources and Siting   89 . crop fertilization. Develop university training programs. SOx.▪▪ Flue gas or other industrial source CO2 supply ▪▪ Investigate the safety of ground-level flue gas logistics. and other sites. and absorption/biofixation efficiency emissions from ponds including plume modeling and algal biomass productivity given seasonal and and regulatory analysis diel variations in photosynthesis and various ▪▪ Effects of various flue gases on algae production water chemistries and co-product quality ▪▪ Control of algal biomass quality (ratios of lipids: ▪▪ Scrubbing of flue gas for NOx. and processing and algal cultivation/technology companies prior to fuel extraction.

1136-1144.. Zubieta. Microalgal biomass for greenhouse gas reductions: Potential for replacement of fossil-fuels and animal feeds. Krause.. CO: U. 150. Biosorption of heavy metals by microalgae in batch and continuous systems.J.. Patin. Bracco.. R. Roadmap for agriculture biomass feedstock supply in the United States.epa. (2009). Japan. DC. Dennehy. Buck. W. Sheehan. (2002). Synergistic relationships in algal- bacterial microcosms for the treatment of aromatic pollutants. 86(3).gov/energy-water/docs/121- RptToCongress-EWwEIAcomments-FINAL. Sokhansanj. B. Environmental Protection Agency (EPA). H. Available online at: http://www.J. Hatti-Kaul. DC.. J. 117–125. Munoz.H. H.html Environmental Protection Agency (EPA). Washington. Hettenhaus. In Water Desalination and Reuse Strategies for New Hess. seasonal. Water rush: Deep aquifers sought for metro growth. DC.sandia. Tam (Eds. J. Lundquist.E.C. and Benemann. Department of Energy.... Clark. D. and Oswald. Available online at: www. J.L.E.. Van Olst.pdf....orau. Energy Information Administration (EIA).S. Emissions of greenhouse gases report.html Environmental Protection Agency (EPA). Low cost reclamation using the Advanced Integrated Wastewater Pond Systems® Technology and reverse osmosis. Adey..B. (2004). (DOE/EIA-0484). (1982). T. Alley. International energy outlook 2009 (chap. Brune. L. S. Clean Watersheds Needs Survey 2004 Report to Congress.Y. Naucler.pdf Department of Energy (DOE).J.L. New Mexico Water Resources Research Institute.. Available online at: http://www. Springer-Verlag.References Aksu. Jessup.R.C... 90 9. (2009. (2006b). A. September). B.). A.. (2009)..R. S. K.doe. J. DC. 8). Easterly.. 2009). Washginton. (2006a). (NOAA Technical Report NWS 34). W. (2008).C. Kyoto. and Ruijs.. B..L. J. (2003).. Ocean Coastal Management. 2002. Journal of Environmental Engineering. H. Foust.. Turhollow.pdf Downing. greenhouse gas emissions and sinks: 1990–2007. (USGS Fact Sheet 075-03). Available online at: http://genomicsgtl. Washington. F. Washington. Bioresource Technology.. R. 189-203. X. U.R. J. Ku. Washington. Resources and Siting   .K. Washington.. R. J. 45(1). and Oswald. W... Delgado. Human-related sources and sinks of carbon Farnsworth. (DOE/SC-0095). D. 95-122. Carbon capture & storage: Assessing the Economics. D. and annual pan evaporation for the United States. October. Guieysse. DC. Department of Energy.B. (1998).htm Craggs. (February 11. In Y-S Won & N. April 15).. The controlled eutrophication process: Using microalgae for CO2 utilization and agricultural fertilizer recycling. I.R. Evaporation rate as a function of water salinity. Available online at at: www. Erbach. Massingill. Desalination. (2008). and Thompson. Hoskinson.Y. M. Denver.J. Environmental Protection Agency. 293-300. Geological Survey. Energy Information Administration (EIA). J..abqjournal. (2002). Wastewater Treatment with Algae (37-54). Breaking the biological barriers to cellulosic ethanol: A joint research agenda. (2008). J. Washington. Berlin.K. Available online at: http://www.S. Al-Shammiri.F. G. W. Energy demands on water resources: Report to congress on the interdependency of energy and water. Extensive open ocean aquaculture development within wind farms in Germany: The prospect of offshore co-management and legal constraints. and Mattiasson. National perspective on saline aquifers. A. O.R. R.F. W. Environmental Protection Agency. Department of Energy (DOE).. Weissman. Inventory of U. 135(11). Albuquerque and Brune. Mean monthly. M. 47. Desalination of groundwater: Earth Science Perspective.J. Available online at: http://www. Borde. Lundquist. H. DC.eia... A controlled stream mesocosm for tertiary treatment of sewage. Thompson. (2003).X.. (DOE/ EIA-0573). Cushman. Available online at: http://www.N. T. DC. (2002). 149-169. D. Environmental Protection Agency. T. (2002). (2007). Green.. B.. Campbell. J. Water Science and Technology. J. Benemann. US Department of Energy. Tagore. (2004. US Department of energy Office of Science and Office of Energy Efficiency and Renewable Enery.J.J. (1996). Ecological Engineering. McKinesey Climate Change Initiative. D.doe. R. E. and Rosenthal. and Wright.. Department of Energy.

natcarb. Cameron.. energy. 12). E. (2003).. D. Treatment of dairy manure effluent using freshwater algae: Algal productivity and recovery of manure nutrients using pilot-scale algal turf scrubbers. H. and Midgley...T. Effects of salinity upon evaporation from pans and shallow lakes near the dead sea. Geological Survey Hydrologic Investigations.E. Maxwell. Feinberg... (USGS Circular 1268). M and Pierce. and Hogg. McNeill. and Meyer. I. 452. U. National Academy of Kohn. Solar Energy Research Institute. (2005). Modelling the water and salt balance in a shallow lake. and Terry. ORNL/TM-2005/66). Sandia National Laboratories. and Karpuk.R.CO2 sources were compiled by NATCARB http://www. Hsieh. University of Illinois. Development of Drainage Treatment for the San Joaquin River Water Quality Improvement Project. B.F. M. M. Ahmadzadeh B.W. The energy challenge: Commentary. T..natcarb. (1978). and the environment..M. Water implications of biofuels production in the United States. D. (1988). Hydrological Processes. Cambridge University Press. Impact of chemistry on sustainable water with its nexus to the economy. Green. K. Hightower. D.. In Metz.C. Carbon Dioxide Capture and Storage. 52. Resource evaluation and site selection for microalgae production systems. and Donald C. Pate. Meyer. Oroud. In Borowitzka and Borowitzka (Eds. Anthony F. Saline aquifer data http://www.. Hill. S. R. Huang. Ecological Modelling. United States Geological Survey. 231-240. Folger. R. UK.A. Grandbois. L. M.J. Hightower. Kokya. S.. CO2 sources for microalgae-based liquid fuel production. Lundquist. O.C. Linsey. 2005-2012. Guieysse. 22. R.K.S. Resources and Siting   91 ... (OPEN-FILE REPORT 6-2003). Evaporation atlas for the contiguous 48 United States. Oswald. Pizarro. Solar Energy Research Institute.. 41(13). C. Denver.B. National Academy of Sciences (NAS). E.A.. S. M. Feth.F. Oklahoma Geological Survey.. and Mattiasson.. (2008).. Golden.. Kenny. Proposing a formula for evaporation measurement from salt water resources.. I. and Maupin M. (2008). N.S.. and Oswald.S. CO. M.C. 51(12). (2009). OK. K..).html NATCARB (2008b).S. (2007). Microalgae and wastewater treatment. N. NM. T.. Turhollow. Department of Energy. The National Academies Press. Microalgal Biotechnology (chap.. 4. S. H. Golden. Lumia. Barber. Environmental Science & Technology. Kenny. Water Science Technology.K. Wright. CO2 Source Map of US .. Robert D. C.L. Reynolds. Fuels from microalgae: Technology status. Theoretical and Applied Climatology. Fresno. Washington. Rolvering. Los Alamos. Hutchison. Available online at: http://pubs. J.L. Norman. U. (SAND 2007-1349C).S. Mulbry.. Overview of energy-water interrdependencies and the emerging energy demands on water resources. B. and Peck. (1986).. Solar Energy Research Institute.. California Department of Water Resources.Y. and research requirements.. C.. W. and Kebede-Westhead. (1995). Davidson. 4626-4632. (1990). Biomass as feedstock for a bioenergy and bioproducts industry: The technical feasibility of a billion-ton annual supply. 285-286. A.G. (1985). (2007). Estimated use of water in the United States in 2005. 211-235. F. Linsey. E... R. 8137-8142. (2004). A. Erbach (2005). W. Thompson.Farnsworth.S.. de Connick. 9. Center of Advanced Materials for the Purification of Water with Systems. (2008).P. Association with natural organic matter enhances the sunlight-mediated inactivation of MS2 coliphage by singlet oxygen. K.K. D. (SERI/SP-231-2550).. B. Perlack. Shannon. S. C. B. and de Coninck. E. Department of Agriculture. Produced water and associated issues.. (SERI/TR- 215-2484). Stokes. California..html Neehan. Lovelace. DC. (Atlas HA-199). (1982). Preliminary map of the conterminous United States showing depth to and quality of shallowest ground water containing more than 1. L. J. (NOAA Technical Report NWS 33) Washington.232-3820). Bryce J..H. (1965).. Barber. Golden. Loos. CO. Estimated Use of Water in the United States in 2000. Muñoz. and Einfeld.. 261-265. K. N. Cambridge University Press. W.000 parts per million dissolved solids. potential.E. Feinberg. Washington. J. and Nelson.L. M. J. Graham. Technical summary. Photosynthetically oxygenated acetonitrile biodegradation by an algal- bacterial microcosm: A pilot-scale study. CO. R. (2005). (Eds). Lynn L. Bioresource Technology. CO.L. (2004). Quinn. Rubin. and Maupin.A. (DOE/GO-102005-2135. Borglin.G. T. Nature.. and Kokya. (USGS Circular 1334). S. E.. Hutson.. (2007).usgs. (2006). Hutson.L. NATCARB (2008a). (SERI/TP. D.S.J. R. Kondrad. Robin L.J. 99. K. McIntosh.

J..A.R. and Perlman. Pierce.1122-1131. Benemann. Journal of Environmental Engineering. Department of Agriculture (USDA). CO. P.Sheehan. (ANL/ ESD/09-1). M. National Renewable Energy Laboratory. Major uses of land in the United States. Geological Survey. Solley. R. Wu. Produced Water Quality Map of US http://energy.A. T. I. D. Golden. S. Applied Environmental Microbiology. Lynch. (2009).S. Darzins. (1998). Geological Survey Circular 1200). D. 92 9. 246-250. 135(11).htm U. W. Woolhiser. and management: Major uses of land. Weyer. (1993). P.ers. In 68(3). 4). Available online at: http://water.. C.G.htm U. Land use.. R. D. 1993. K.S.R. R. Sinton. Resources and Siting   . Wang. J.. Estimated use of water in the United States in 1995. value. (Economic Information Bulletin Number 14).S. D. Department of Agriculture (USDA).)..C. Department of Energy’s aquatic species program--biodiesel from algae. M.S. Sunlight inactivation of fecal indicator bacteria and bacteriophages from waste stabilization pond effluent in fresh and saline waters. and Wallace.J. (2009). and Davies-Colley. B. McGraw-Hill... USDA Economic Research Service On-Line Briefing Room Available online at: http://www. (2009).W. Geological Survey.. W. and Nelson. Mapping average daily pan evaporation. A.E. 2002. 1115-1122..usgs. M. M. (U. Feffer. 110(2).. A. Theoretical Maximum Algal Oil Production. (2009. Argonne National Laboratory. Mintz. (Ed).usgs.usda.A. & Willson.. Consumptive water use in the production of ethanol and petroleum gasoline. (2009). BioEnergy Research.M. (1984). A look back at the U..J. Lipid productivity of algae grown during treatment of dairy and municipal wastewaters.S.... Y. Hall. and Roessler. (NREL/TP-580-24190).B. U. Handbook of Hydrology (chap. L. Woertz. Journal of Irrigation and Drainage Engineering. (USGS Circular 1331). USDA Economic Research Service. Bush.. (2006). Climate change and water resources management: A federal perspective. Dunahay.H. (1998).S. US Geological Survey.S. T. Shuttleworth. (2002). D. J. (2002). and Arora. 1–10.. Evaporation.

Toward this end. Systems modeling and analyses technical innovations in systems and processes. can commercialization of the still relatively immature provide informed guidance for targeting investments in field of algal biofuels also requires the confidence and select technologies. tradeoffs among different technologies.Separation Hydrocarbons Consolidated Intermediate Lipids Carbohydrates Proteins & Alcohols* Biomass Constituents * Biologically produced in-situ by the organism Biochemical Thermochemical Direct Anaerobic Conversion Conversion Conversion Synthesis Digestion Processes Renewable End Use Fuels Biodiesel Alcohols Biogas Co-Products & Products Hydrocarbons 10.Dewatering . systems. as discussed co-products industry requires the optimum combination of throughout the Roadmap. and environmental) critical technical risks and overcome challenges to developing to meeting performance objectives and reaching goals for an algal biofuels industry. and deployment engagement of key public and private stakeholders so approaches. It can also help track performance and they can make necessary investments over time to reduce progress (technical. significant investment is expected to be required to overcome the various technical and economic Exhibit 10. Such serve a range of purposes and goals that include: analyses can provide improved understanding and insight to help guide successful industry development within the • Assess technical performance and cost/benefit real-world context of technical. as illustrated in Exhibit 10. Systems and Techno-Economic Analysis Successful development of an algae-based biofuels and challenges along each of these pathways. and market conditions and constraints.g.1). e. economic.. techno-economic modeling and analysis can across the overall algae-to-biofuels supply chain.10. Systems and Techno-Economic Analysis  93 .Harvesting Systems . systems.1 Objectives of investments. and quantitative modeling and analyses of systems and processes are needed that span different enterprise levels In short. infrastructural. comparative yield and energy balance for different pathways • Assess economic impact of R&D strategies and 10. and processes. ranging from individual unit and integrated scale-up for commercial production operations to an integrated enterprise. to an overall regional and marketing.Fractionation Phototrophic Mixotrophic Phototrophic Heterotrophic Phototrophic .2. objective successful commercialization. processes..g.1 High level illustration of various approaches and pathways to developing algae-derived biofuelsand co-products ALGAE Species Unicellular Multicellular Selection & Cyanobacteria Microalgae Macroalgae Biology Open Terrestrial Closed Terrestrial Open Offshore Cultivation Systems Systems Systems System: Hybrid . environmental. political. e. comparative cost per Techno-Economic Analysis gallon of biofuel for different pathways Given the numerous potential approaches and pathways available for algae biofuels and co-products (as illustrated in Exhibit 10. coupled applied across this space and done at different levels of with economic feasibility in the practical implementation precision and granularity. Enabling successful advancement and or national resource and supply chain assessment.

• Assess environmental impact of R&D strategies 10.2 TEA Scope and Methods
and investments, e.g., comparative net greenhouse
gas (GHG) emissions per gallon of biofuel for Exhibit 10.3 illustrates at a high level the essential
different pathways elements of techno-economic modeling and analysis.
• Assess consequences and constraints of alternative These intersecting elements present a broad systems
pathways for algal feedstock, biofuels, and co- perspective that integrates the interdependent science
products industry build-up, e.g., land, water, CO2, and engineering aspects specific to developing algae
other nutrients, energy, and infrastructure. biofuels, keeping environmental, economic, and
policy aspects in view to provide critical insight
• Inform R&D and business development investment
and information needed for decision-support.
• Inform policy decisions and explore “what if” Techno-economic assessment for algae biofuels is
scenarios highly interdisciplinary and will range from GIS-based
• Provide insight and actionable information needed resource and siting assessment, to engineering modeling
to successfully guide technical advancement, of systems and processes at the unit operations level, to
integrated system scale-up, and commercialization interdependency analyses of the entire supply chain. Exhibit
10.4 illustrates a broad systems perspective of issues and
DOE’s Biomass Program employs a wide range of approaches for the overall algae biofuels supply chain from
analytical tools, data, and methodologies to support siting and resources to product end-use. The categories
decision-making, guide research, and demonstrate shown follow the process steps in the algal biofuel supply
progress toward goals. In conjunction with the Algae chain sequence. Such summarization of key areas to be
Roadmap, Biomass Program’s strategic and portfolio addressed to enable algal biofuel production provides
analysis efforts are expanding to include algae with guidance for the scope of content that should be integrated
techno-economic modeling and analysis (TEA), life cycle into systems techno-economic modeling and analysis.
assessment (LCA), and geographic information system
(GIS) based resource assessments.

Exhibit 10.2 Systems and Techno-Economic Modeling and Analysis as a decision-support tool
It can track and assess performance, and evaluate investment cost/benefit/risk tradeoffs to inform both technical and programmatic decisions

S&T, Systems & Processes Areas
Biofuel Conversion
Cost, Performance, and Risk Metrics

Extraction & Fractionation
Harvesting & Dewatering
Algae Science & Biomass Production
Stage-I Advances Systems Infrastructure & Integration
Uncertainty Range

Cost & Risk
Reductions Stage-II Advances
Improvements Commercialization Threshold

Cost & Risk

T0 (current status) T1 T2 (e.g., 5-15 years)

Roadmap Timeframe Multi-Stage R&D,
Demonstration, Commercializtion, Deployment

94 10. Systems and Techno-Economic Analysis 

Energy & Fuel Economics; Algal Biomass & Biofuel
Energy-Water-Environment Production Systems,
Interdepencies; Process Engineering
Carbon Emissions & & Other Sciences

Policy Analysis

Exhibit 10.3 Techno-Economic Modeling and
Analysis addresses interdependent issues

The scope of TEA will ideally include the capability to level, which in turn must function within climate and
apply and integrate detailed process engineering and weather conditions that vary with geographical location,
physics-based modeling at the unit operations level the approach needs to be both multi-level and multi-scale.
with multi-scale and multi-path systems modeling and
comparative tradeoff assessment. This can include An algae biofuels and co-products supply chain will be a
geospatial (GIS-based) data collection, analysis and complex interdependent system with numerous alternative
visualization, systems and processes optimization, pathways and functional elements and feedbacks at various
and dynamic interdependency analyses of the overall spatial and temporal scales and resolutions, as suggested
supply chain at a local, regional, national, or global in Exhibit 10.5. System and process simulation and
scale. A major environmental impact element will optimization under a system engineering framework can
be life cycle analysis (LCA) to assess GHG impacts prove very beneficial for system design and operation. For
and other resource use footprint (e.g., water). example, optimizing conditions that produce the highest
neutral lipid content of the algal biomass under a range
The overall goal is to capitalize on the power and insight of design conditions, when bio-oil feedstock production
available through application of well developed computer is the primary pathway and objective, may not lead to the
modeling and analysis tools combined with disparate highest overall biomass or lipid productivity. Similarly,
database information that exists or can be developed. achieving the most affordable and sustainable production
For example, a process-based algae growth model that of algal feedstock, in terms of resource use intensity
accurately reflects and integrates the dynamic behavior of and cost per unit of end-use biofuel produced under a
the key biological processes with the engineered growth specific set of scenario conditions, may not result from the
system and its time-dependent physical and chemical pursuit of the algal oil production pathway altogether.
conditions (including local time-varying climate and Sensitivity analyses and global system optimization
weather) can not only provide more detailed input to and comparative tradeoff assessments across a range of
the techno-economical performance analysis of algae approaches and conditions are among the critical modeling
cultivation, but can also reveal opportunities for improved and analysis needs. Meeting such needs requires a flexible
system and process design and operation. Since the modeling and analysis framework and suite of tools that
characteristics of the biological system at the microscopic can specifically address the broad scope and multiple
algae cell level affect the performance of the engineered pathways spanned by an algae biofuels supply chain
cultivation system and processes at a higher integrated system. Geospatial GIS-based data integration and analysis

10. Systems and Techno-Economic Analysis  95

Exhibit 10.4 Scoping the content for Systems Techno-Economic Modeling and Analysis: Key topic areas and issues for the overall algal
biofuels supply chain


Algal Biology Cultivation Harvesting & Extraction & Processes Biofuels Policy &
Sitting & Inputs
S&T Systems Dewatering Fractionation Co-Products, Regulation
& Services

Geolocation/ Species Selection Open Ponds Filtering Extract Processes Conversion Processes Taxes
elevation & Matching to - lined Flocculation/ - Solvent - Biochemical Incentives
Land characteristics Growth Conditions - unlined Settling - Acoustic - Thermochemical
- Electromagnetic - Digestion Permitting
Characterization - raceway
Climate/Weather Airlift Flocculation - Other - Hydrotreat/ Enviromental
- wastewater
Solar Insolation Performance treatment Centriuge Seperation/ Refine Impact
Water Souces/ Strain Improvement Closed PBRs Drying Fractionation Fuels Health & Safety
Issuses - horizontal tube Biological Assist - Membrane - Biodiesel Algae Control
- brackish Biomass Growth
- vertical tube - brine shrimp - Distillation - Green diesel & Regulation
- wastewater & Oil Conditions
- vertical planar - fish - Centrifuge - Aviation
- produced Photoautotrophic Other
- other - Other - Gasoline-like
- desalination organism operation Other
Hybrid System Intermediate - EtOH
concentration Products - Biogas/methane
Heterotrophic Combinations
-marine - TAG Oil - Other
organism operation - covered ponds
- fresh - Other Lipids Co-Products
Algae Pathogens, - pond/PBR mix
- losses, re-use ▪ Polar - Feed
- salt build-up Predators, and Heterotrophic ▪ Neutral - Fertilizer
CO2 Sources Mitigations - Industrial - Carbohydrates - Chemicals
- power plants Bioreactors - Proteins Services
- cement plants - Wastewater - Other - Carbon capture
- fermentation/ Facilities Compounds - Water treatment
other - Water
Chemical/Materials Monitoring & Direct secretion
Energy/Power Maintenance of EtOH or
hydrocarbon fuel
precursors into
growth medium,
avoiding Harvest &
Dewatering steps

is also key to siting and resource assessment and the design, uncertainty in data while still capturing the underlying
analysis and optimization of facilities location and supply dymanics of the overall system. Although quantitative
chain logistics (from algae cultivation through end-use uncertainty will remain, the results can provide qualitative
fuels and products biorefining, transport, and distribution). system behavior and trends still useful in helping
guide technical, economic, and policy decisions. More
Algal biofuels remain an emerging field at a relatively thorough analysis and modeling refinement, informed
immature stage of development. As a result, data for by field data expected to become available as larger-
detailed unit operations and process analyses for different scale pilot and demonstration projects are established
technical approaches for algae cultivation, harvesting, and and operated, will more clearly reveal the most critical
post-processing into fuels and other co-products is currently challenges and guide investments and technical advances
sparse and uncertain. Selected modeling and analysis needed to enable successful commercialization of
approaches can be used to effectively accommodate such economical, scalable, and sustainable algal biofuels.

96 10. Systems and Techno-Economic Analysis 

Primary Energy Electric Power and Process Heat Generation Capital Construction.Climate/Weather Feedbacks Conversion Carbohydrates Biofuels Algae Cultivation .Requirements Harvesting Dewatering Extraction Oil / Lipids . individualized modeling and analysis efforts and configurations of rapidly evolving systems and by industry and technology development organizations processes. It was noted that there Preparation for the Algae Roadmap Workshop and the is a need and role for both integrated systems modeling as discussion among participants during and after the event well as detailed engineering design and process modeling.CO2 / Flue Gas Conditioning Other Coproducts . Workshop participants the role that systems TEA and LCA can and should play generally agreed that modeling and analysis should be a in support of algal biofuels development.Land (cost.Solar Insolation Environment . Incentives & Barriers Siting .6 illustrates various complementary modeling and scales will be defined in the early stages of the systems analysis approaches and techniques that come into play at modeling R&D effort and will evolve as time goes on.Materials Energy Feedbacks . One approach with photoautotrophic microalgae integrated systems modeling and analysis across the is modeling major cultivation system categories 10. identified the need to clearly define the scope and determine and that the two should be coupled. be considered in algae biofuels modeling and analysis production plant. Demand & Price for Co-Products vs. Included in the Workshop discussions was the question of Due to their more limited and immediate business how best to approach the multiple and disparate paths objectives. as exemplified by Exhibit 10.Equipment . similar to the modeling approaches. scales.GMOs Biology . Policy & Regulation. soil) Broader .Nutrients (NPK) or Treatment & Re-Use . tilt. Maintenance.5 The dynamic coupling and interdependencies across the algal biofuels and co-products supply chain 10.Characteristics . or enterprise level rather than broader efforts.Water Water Capture . develop. Cost of Petroleum & Conventional Fuels.Autotrophic . Details of systems’ and processes’ technical and economic performance achieved by industry and LCA Approach (Workshop other technology developers are also usually considered Results and Discussion) proprietary. Also identified critical part of a national algae biofuels research program was the need to adapt.Labor . etc.Closed systems &/or Drying & Seperation . It is also expected available and emerging information and data.Performance .7. unit operation.Open systems . that should tend to focus on a technology or process. geology. unavailable for broader use and objective assessment and validation. location. Exhibit 10.Heterotrophic . different levels within the broad systems scope presented.Species . Operations. Monitoring. their alternatives. and therefore.3 Suggestions for TEA and supply chain. Systems and Techno-Economic Analysis  97 . Exhibit that the needs for coupled models of differing fidelity and 10.Hybrid systems Feedbacks Resources Proteins & Conversion .Temperatures Design & Economy Feedbacks Nutrient . and levels of detail best suited to and analysis efforts underway by DOE in support of the perform the necessary modeling and analyses based on other bioenergy and biofuels programs. Replacement Market Externalities: Cost of Energy. and utilize a range of and industry development effort.Capital Co-Gen .

across an emerging approach.Integration with existing infrastructure . GIS Analysis. 98 10. platform). production availability. and policy decisions to best conceptualize a model that spans the overall at the state. with time delays.Vendor/ Supplier cost estimates (ASPEN+) • Engineering and construction cost estimates • Process GHG footprint (LCA) assessment . specific technologies. and national level. economic.g.... transport. and Life Cycle Analysis of open pond. The disadvantage of such a be established for system and process functional blocks that distributed and narrowly focused approach is that it is often would enable the development of an open-source modeling done in isolation with limited or no consideration of other and assessment platform with “plug & play” flexibility. • Other infrastructure and enviromental features Exhibit 10. or a hybrid competing options and the broader scope of interdependent combination. enviromental. with learning curves & improvement projections .ICARUS cost estimate software (or equivalent) • Operations & Maintenance cost estimates Geographic Information System (GIS) Analysis and Visualization • Cost (& offsets) of co-product feedstock • Land resources (characteristics. storage • GREET (Argonne National Lab) infrastructure • LEM (UC/ Davis). economic. constraints & impacts . Flexibility and usefulness Developing numerous detailed models that are each could be increased to the extent that such modeling uniquely customized to a specific combination of systems and analysis can be done using common standards. Etc. Narrowly “beginning-to-end value chain” for algal biofuels focused and detailed technical and economic modeling production with adequate resolution and detail is very and analysis is valuable and necessary for those doing challenging and still emerging.Engineering & construction estimates from industry . and essentially represents what exists today with “community of practice” for the algae biofuels industry. and interface approaches. how better inform technical.. processes and integrated systems for commercialization.6 Multiple levels and complementary approaches available for Algal Biofuels Systems and Processes Techno-Economic Modeling.. and optimization of of possibilities could quickly become unmanageable. Higher-Level Dynamic Meta-Systems Modeling (Integrated Analysis Framework) • Broad value-chain scope. from resources & sitting through production to end use • Algae biofuel and co-products industry scale-up potential. resource use. and processes and performance parameters is another definitions. can link to other models & analyses Process Flow Enginering Modeling Static CAPEX & OPEX Calculations • System and process equipment cost estimates • Mass & energy balance calculations . pipelines) Other LCA Modeling • Fuel processing. development. • Cost (& offsets) of co-products production other) • Carbon footprint cost accounting • Climate/ Weather/ Temperature Conditions • CO2 resources ( point source emitters.. and activating only those portions of the issues and tradeoffs that need to be addressed together to hybrid configuration model desired. The size of the “matrix” engineering design. and policy issues • Feedbacks and Multiple Sector Interdependencies...Required build-up of new infrastructure. various groups doing ad-hoc modeling and assessment specifically focused on their chosen approach and Standardized interface requirements and definitions could technologies (e..Technical. regional..) • Cost of biofuel production • Water resources (fresh wastewater. Systems and Techno-Economic Analysis  . closed photobioreactors. etc. Beyond this.

USDA.3 Centrifuging biomass 3. computational fluid dynamics and analysis platform that can be productively used modeling of open raceway ponds or closed photobioreactor and leveraged for algae biofuels development. and represents utilizing different techniques such as high performance an example of a GIS-based information gathering physics-based modeling (e. Materials 2.1 Fuel 5. Biogas / Methane Metabolites I.0 Harvesting L.3 CO2 2.4 Off-shore systems 3. Carbohydrates O.Distribution . Fatty Acids M.1. Feed 1.g. Hydrogen K.1 Microalgae Conservation F. etc.2 Open system 2.0 Extraction Conservation 6. A system is an aggregation to different stakeholder communities such as: of subsystems interacting such that the system is able to deliver an over-arching functionality.5 Biological Assist 2. processes.) 4. Gasoline-like 6.3 Chemical G. Dried Algal 1.0 1. cultivation systems) or process engineering models using widely accepted and applied commercial process modeling tools like AspenPlusTM or customized spreadsheets.2 Fuel .1. a process flow diagram was economic/policy R&D and assessment developed and presented at every breakout track discussion to illustrate the intricate interdependencies of algal • Industry developing and commercializing technologies. Ethanol 1.1 Solar .2 Electromagnetic 5. Systems and Techno-Economic Analysis  99 .Field .5 provides • DOE & national labs doing R&D. 10.2 Co-products 1.3 Hybrid systems & settling 1.1 Physical (Thermo/Mechanical) 5. 4.2 Cyanobacteria R.4 BY PASS A.) and non-technical issues.1 Algae species 3.0 J.End Use Exhibit 10.2 Filtering 8. biofuel production.5 Light or Alt.3 Biochemical E.1.Transport fish excrement.3 Macroalgae R.0 Drying . Triglycerides N.1 Flocculation 2. moving in this direction (Getman. and a high-level illustration of the interdependent character tracking of program investments of the overall algae biofuels value chain that involves a • Other federal and state agencies (Department of broad range of systems. 2009).4 BY PASS 6. universities.1.5 Biological assist Harvesting (shrimp. Lipids H. 5. Chemicals 2. and national labs.Storage 4. etc. Energy Source derivative biomass & Markets 3.7 is a revised adaptation systems. assessment. Proteins P..4 Systems Analysis Flexibility in being able to link custom subsystem or process models into an overall meta-system Overview modeling and analysis framework would provide a System analysis is foundational to designing a strategy capability that could be of significant value and benefit for algal biofuel deployment.2 Thermochemical & Separation D. Wet algal or biological assist Infrastructure 1. Exhibit 10.7 High-level multi-pathway algae biofuel process flow diagram for the algal biofuels and co-products supply chain Much more detailed or custom models of individual Work currently underway on the Knowledge Discovery subsystem or process blocks could then be developed by Framework (KDF) by DOE’s Biomass Program is various others in industry. Other Q.1. Exhibit 10. Aviation 6.3 Water 3. Biodiesel 5. EPA. To facilitate system-level • Universities doing a wide range of technical/ thinking during the Workshop.2 Land Cultivation 3. Fertilizer Inputs 1.1 Closed system 1.1.1 Chemical C. processes of the flow diagram that shows a representative number • Private investment / funding sources 10. and other technical Defense. Green diesel 6.

GIS Assessments sustainable production of microalgae biomass. The impact that availability and cost biological. solar insolation. regional. and organic molecule reforming. evaluate costs. etc. Exhibit 10. wind electrolysis. Systems analysis can help manage the complexity of • Climatic characteristics (temperature. This program could serve as a guide for moving Geographic Information System (GIS) Visualization forward with analysis of algal biofuel production.of multiple path process options available for every step technical information and data about the systems and in the algal biofuel production chain. P. especially where Both Exhibit 10. The cost include AspenPlusTM and FLUENTTM. The overall viability and examine the sensitivity of different options for hydrogen production include goal gasification. and comparing trade. H2A was designed to consider various highlighting areas for research and development. nuclear energy. These Engineering analyses (technical and economic) at the include climate and the adequate availability of water. Example engineering analysis tools scale-up was touched on earlier in chapter 9. Inputs to each will likely emerge. A substantial number of barriers are (characteristics. • Fuel processing. among others. identifying and appropriately modeling • Water evaporation loss (function of climate. and others beyond balance systems level view of an algal biomass and algal those selected for inclusion in the figures also exist or lipid cultivation and extraction system. and each numbered node attempt to develop a comparative analysis based on so represents a mass or energy balance calculation. development of mass and energy based systems models energy and environmental tradeoffs and set research can help evaluate different proposed processes for priorities and inform policy by sound analysis. availability. This may also require coupling processes that made up different thematic topic discussion with appropriate external data and driving functions sessions in the Workshop are all inter-related. • Other infrastructure and environmental features Engineering Analyses and Several critical resource factors will impact large-scale. assumptions in individual processes to the overall system. The many process permutations. Critical climatic and natural (energy requirements and costs) of the systems and resource data can be readily accessed. from algae growth processes involved to appropriately formulate and conduct to fuel and co-product processing and end-use.8. The pathways toward a hydrogen economy.5 and Exhibit 10. etc. number of permutations of potential pathways to algal biofuel production.8 represents a mass and energy on the product or co-product chosen. thus Fuel Initiative. and understanding key information from each node in the and other local environmental conditions. Established in 2003 in response to the Hydrogen what steps in the process are most energy intensive. • CO2 resources (point source emitters. characteristics of algae themselves and the characteristics and national scales.8 represent a large detailed process engineering data may be lacking.8. unit operations level require the systematic calculation efficiency of water use. Simpler and process becomes the primary task of the systems analysis. Though it may seem daunting to system component are shown. and benefit tradeoffs of CO2 capture through biofixation which require sufficient understanding and detailed using photoautotrophic microalgae cultivation enhanced 100 10. producing algal biofuels and co-products by quantifying precipitation. hydrodynamic.) uncertainties. Many of the process steps will differ depending Exhibit 10. etc. and Analysis tools (referenced also in chapter 9) are indispensible for algae production and supply Other chapters of this Roadmap point out the lack chain systems analysis due to their ability to perform of availability of detailed information about the mapping and resource analysis spanning local. Collecting such as time-dependent solar insolation. such as: processes that are shown in the process flow diagram • Land and water resources of Exhibit 10. risk. there is precedence for this value of this kind of systems mass and energy balance sort of undertaking in DOE’s Hydrogen Analysis (H2A) assessment is that it can help assess the overall viability Project within the overall Hydrogen Program (DOE. availability of suitable land.) enumerated and designated as goals to be achieved. K) supplies. and environmental impacts. Sub-level accurate analyses. of a given algal biomass production system and show 2009). more approximate preliminary spreadsheet analyses are more appropriate as a first step. and tracking of mass and energy balances that include and availability of supplemental CO2 and other nutrient evaluation of the thermodynamic. (N.) interdependencies and feedbacks. transport. pipelines) offs from various scenarios with regard to cost. storage infrastructure technical performance. Systems and Techno-Economic Analysis  . and chemical kinetics of the systems and of these resource can have on algae biofuel production processes used. most of which are still immature and An example of this type of analysis is presented in emerging. temperature.

The conditions with multiple species of algae could be useful. geographic location attributes and time dependent climate The economics of algae-based biofuels and co-products and weather data. the data available to implement this type of analysis are thought to be preliminary and refinement Producing biofuels that are highly compatible.g. Potential facilities relative to upstream algae biomass production CO2 capture and utilization in algae biomass production and downstream fuel and co-product markets. sourcing. e. etc.. Land. and biorefining into fuels and (e. Equipment. Operation & Maintenance cost estimates transport. is discussed in more detail through a few selected The combination of GIS resource assessment.. The possibility fuel and co-product markets at the end of the suppy chain.. or totally based on model improvement and validation with data fungible.g. re-use Potrential Protein Biofuel Feedstock Materials. P. Co-products Polar Lipid Site Preparation Carbohydrate 10. Hexane microfiltration e. can provide powerful feedstock production and downstream processing can be capabilities for estimating performance and for improving integrated with existing infrastructure and markets. with physics-based modeling of will depend in large measure on the degree to which algae cultivation systems and algae growth. Midstream will be the includes an algae growth model driven by environmental logistics of transport. followed downstream by the necessary concentrations.. Exhibit 10.g. and Capture & Neutral Lipid Systems Infrastructure. Primary Secondary Primary Secondary Neutral Lipid Bioreactor Harvesting Harvesting Dewatering Solvent Air & CO2 Dewatering Extraction &/ or Pond e. Structures. the economics can be expected to improve. the addition of CO2 from industrial sources will affect on the size and the optimum site locations of biorefinery the economics of algal biofuel production. etc. and energy). supply chain. and transport of water.) Water Energy Residues: TAGs & Other Facilities. storage. including example calculations presented later in this chapter. Hexane System Chemical Dissolved Air Recovery & Centrifuge Drying Solvent Nutrients Flocculation Flotation (N. Currently. in press). nutrient other co-products.g... storage. Systems and Techno-Economic Analysis  101 . same would apply to co-products and their potential markets. from the operation and monitoring of larger scale algae distribution.) forcing functions and boundary transport. and distribution into the various end-use conditions (James and Varun. K. the relative location and • ICARUS cost estimate software (or equivalent) logistical costs associated with fuel processing. analysis and LCA (CAPEX ) and operational expense (OPEX) calculations for algae. • POLYSYS At the downstream end. Biomass e. with the appropriate models.g. tools and capabilities to an open raceway pond system that supplemental CO2. and distribution infrastructure could benefit from GIS analysis. for • Discounted cash flow analysis example. ambient temperature. and policy decisions.8 Notional baseline algae feedstock production system Sunlight H2O Energy & Solvent & Water & O2 Other Inputs Energy Energy H2O Recycle Microalgae Photo. tools. this will include the logistics different site conditions. of combining these types of spatial and physical data to To the extent that operations can be effectively co-located project possible algal production scenarios as a function of or established within reasonable distances across the geography and climate is an interesting avenue of inquiry. These and related issues should be an integral Economic analysis tools for static capital expense part of techno-economic modeling. with the existing hydrocarbon fuel handling. e. and data are also integral to system analysis as they reveal financial sets developed and leveraged to provide the necessary investment or market incentives needed for algae biofuel assessment capabilities to guide research.9 is an example of the and costs of supplying inputs needed for algae production application of high-performance computational modeling (suitable siting. solar insolation. This information. At the the technical design of systems and their operations for front end of the supply chain. e. Utilities. • Equipment.. resource management. storage. business deployment.g. can help determine the practical upper limit • Cost (& offsets) of co-product feedstock production • Cost of biofuel production • Carbon footprint cost accounting Exhibit 10. Some examples are: development.g. and end-use infrastructure would result in production systems under a range of site and climate easier and more widespread market acceptance.

on the other hand. climate. aspects of large scale agriculture. availability. vary over the U.) will also affect both conditions. but will variation of resource availability will have major impacts be dependent on the geographical location. are amount of water required nationally could begin to not unique to the development of algal biofuel technology.. and sustainability of suitable water Impact of Geographic Variability of supplies for algae production will be a key input factor Inputs on Algal Biofuel Production Costs for inland cultivation.700 gal/ac-yr. P. 50% oil content. can potentially help fill this need. productivity. and it will affect the amount limit on the order of ~4. and this factor varies widely across the Sugar from commodity crops and other organic country among inland. but these commercial algae industry build-up scenarios. wind. etc. Si. and fertilizer. and annual variation in other assumptions made in the analysis (which include solar insolation. seasonal. O2…) to model growth rates. and will be heavily dependent on geographical location and local conditions. precipitation. particularly with open systems subject to macroalgae and microalgae cultivation (autotrophic. Exhibit 10. Capture and re-use of non-fresh water. though important for overall TE analysis. and offshore sites. temperature.9. Areas The various inputs necessary for algal biofuel production of the country with the highest solar resource best have been described earlier in this chaper and in other suited for algae growth also tend to be more arid and chapters of the Roadmap. and nutrient gradients • Solar insolation and other environmental. cost. Open raceway pond example of physics-based algae cultivation system modeling and anlaysis. evaporative loss. coastal. Computational Fluid Dynamics Model of Algal Raceway Pond Using Modified EPA and US Army Corps of Engineers Codes • The Environmental Fluid Dynamics Code (EFDC) solves 3D Navier-Stokes equations of open channel flow to model speed. heterotrophic. as well as other climate-related the availability of high solar insolation consistent with factors such as temperature and weather (cloud lower latitudes and/or high percentage of clear weather cover. Such analysis can enable both performance prediction and pond system design optimization after appropriate refinement and validation with monitored field system data (Adapted from James and Varun. and affordable accessibility of such water sources. • Availability. and mixotrophic) for which geographical in particular. 2009). like cost of subject to more limited water supplies.) (Weyer et al.350 . but it is appropriate here to briefly note • The supply. Under large power.000 gal/ac-yr and perhaps a practical that can be captured. and basis. photosynthetic efficiency. it will affect the amount of CO2 the order of ~38. based of culture that will need to be processed on a daily on the expected losses. and cost of organic carbon the following: feedstock needed as input for heterotrophic microalgae production will play a major role in the commercial • The average annual insolation is generally the viability and extent to which national production dominant and rate-limiting factor for autotrophic algal capacity can expand using the heterotrophic approach. carbon materials from industrial or municipal waste This variation will determine the spatial surface streams can provide bridge feedstock in the near- 102 10. approach the same order of magnitude as large scale There are. in press) area of cultivation systems needed to achieve a set Recent analysis suggests an upper theoretical limit on amount of product. the variations.5. water. availability. Systems and Techno-Economic Analysis  . Certain elements. The daily. on cost of production and scale for commercial viability.S. These aspects were discussed in some detail in Chapter 9. etc. the productivity and reliability of production. and meteorological forcing functions • Includes algal biomass growth model • CE-QUAL couples nutrient kinetics and 22 independent variables (N.

. 2009). water evaporation rates will provide some level of and affordable accessibility of such nutrient sources. 1998. and and processing improvements needed to reduce the the associated logistics. and and costs as transporting and distributing CO2. As noted here and in chapter closed systems operating inland can overheat. it may be co-located industries or the CO2 source may allow necessary to transport CO2 over some distance. availability. but this will be systems operating at inland sites in the summer.. if not more. infrastructure facilities for offshore. where recent cost and supply input resources with the required output product issues have had negative impacts on the industry processing and distribution infrastructure and markets. but for each approach operational costs for algae (Massingale et al. 2009) can potentially help supply specific strains for extended durations. If it is agriculture (Huang. the right of access and use. but applying facility will require extensive pipeline infrastructure for this heat or energy to extensive algal cultivation CO2 distribution. and costs of offshore cost of lignocellulosic material deconstruction into marine operations will have a major impact on simple sugars and other organic carbon compounds costs of production. temperature control. scale. transporting. Conversely. Systems and Techno-Economic Analysis  103 . but major sustainable scale-up of national contaminants that inhibit algae growth).e. though it is not yet energy crops. such as are present in some forms of flue gas. lipids. and is a consideration that appropriately processing lignocellulosic biomass requires further investigation. 2005). The affordability of • Land prices and availability can also impact the cost generating organic carbon feedstock from such of biofuel production at inland and coastal sites. (Huang. derived originally from lignocellulosic biomass. Finally. nitrogen than conventional biomass be possible to get the most benefit of economies of crops on a mass balance basis (Alexander et al. The quality of the systems will provide the same engineering problems CO2 source will also play a role for algal growth. the amount of nutrients required nationally versus the matching of affordable production scale to could begin to approach the same order of magnitude the sustainable and affordable supply of the required as large scale agriculture. and municipal waste streams will be location-dependent. adding to the cost. Algae can be production capacity will demand the use of sugars effective at capturing and concentrating heavy metal and other suitable organic carbon source materials contaminants (Aksu. Commercial fertilizer costs are overall cost of total algal biomass and intermediate tied closely to the cost of energy supplies (natural gas feedstock fraction (e. 9. term. This As with cellulosic ethanol.g. 2009.. other) production. In necessary to distribute the facility over a number of the absence of nitrogen fixation. carbon credits materials in the form of woody and herbaceous must also enter into this analysis. on the industrial build-up. but this the cost of operations. and inland • The supply.. availability. N. The capture and reuse of nutrients • As in traditional agriculture. For open nutrients for algae production scale-up. Mehta and Gaur. Even in active thermal management for growth during periods the case of co-location. 2008) it will likely be an optimum minimum and range to the same extent as for large scale commercial of size for a commercial production facility. and cost of other nutrients sites will all be location-dependent.. Cost of site preparation and suitable for feeding heterotrophic microalgae. algae can require as smaller parcels of land or offshore sites. and can be an appreciable factor in proteins. materials will also depend on technical advances For offshore sites. it will be advantageous to co-locate cultivation requiring active cooling that can add prohibitively to facilities with stationary industrial CO2 sources.. K) required as inputs for algae growth will straightforward to calculate the impact of the cost also play a role in commercial viability and extent of of land. It is reasonably (i. Waste heat and energy from will not be feasible in all instances and thus. Malcolm et al. and co-products like animal feed. 2009). but evaporation will also add to • CO2 availability and cost of delivery will play a major operating cost (for water replacement and/or for salt role in autotrophic microalgae cultivation scalability management with brackish or saline water. waste materials from agriculture clear how best to factor this into the calculation. and perhaps also for offshore sites. the size of an autotrophic algae of suboptimal (high or low) temperature. coastal. The key tradeoffs will be between the cost Under large commercial algae industry build-up of overall production (capital and operating costs) scenarios. and petroleum). dependent on the geographical location. carbohydrates. P. and forest industries. some sources are likely to require more cleanup than others (especially if there are plans for animal feed as a co-product and/or if the CO2 source stream includes 10. risks. it may not much. and operating expense. the logistics and costs could impact the suitability of residual biomass for associated with producing. the temperature during from various agricultural and municipal waste streams the growing season will restrict the ability to cultivate (Woertz et al. 2008).

peer-reviewed literature (e. 1993). The process employs a and scenario studies. 2001 and 2002). Delucchi. constraints and impacts 2009). and maintenance. LCA is a systematic technique to assess Systems dynamics modeling is a powerful and flexible the environmental aspects and potential impacts associated modeling approach that can foster collaborative analysis with a product. Bernard. LCA is important for assessing relative Oswald. and other resources. it is system performance for an industry that has yet to be likely that a technology that will require the production commercially realized is one of the biggest challenges for of immense volumes of biofuel at affordable cost so it techno-economic analysis. the calculations for the cost of algal biofuel 10. co-generation addition to net GHG emissions.S. use. waste stream 2002 and 2004). EPA. resource use. white papers / industry analyses. 2004) up systems. economic.5 Algae Production production will benefit from detailed inputs that take into consideration the variations in cost and availability of the Costs and Uncertainties essential elements for cultivation. learning curves and improvement projections • Technical. 2006. 1997). however. 1996). delays. life cycle inventory analysis (LCI). and tools have been developed that include Argonne National • Algae biofuel and co-products industry scale-up Laboratory’s GREET “well-to-wheels” model (ANL.g. EPA. and • Broad value-chain scope: from resources and interpretation as illustrated in Exhibit 10. nutrients. to final disposal. algal oil feedstock Life Cycle Analysis production cost analyses were reviewed based on a Life Cycle Analysis (LCA) is a “cradle-to-grave” range of sources to gauge the status of algal biofuel analysis approach for assessing the resource use and technology. These 1993. that a and including algae (Kadam. energy) impacts among different approaches to personal communications with field experts. UNEP. or service. modeling. eliciting discussion.11 illustrates the typical life cycle stages considered in an LCA and the typical inputs/ Dynamics Modeling outputs measured. and in comparison with fuels citable sources are quite dated. energy. While these variations Data gathering and validation of technical and economic may be minor relative to the technical uncertainties. siting through production to end use LCA methodologies. LCA for biofuels can also • Integration with existing infrastructure assess impacts and tradeoffs associated with utilization • Required build-up of new infrastructure with time intensity for water. from manufacture.. potential. Tapie and GHG emissions and other resource utilization (e. environmental. output flows. Department of Energy (e. process. operating costs. ISO. Benemann and and processes. life cycle impact assessment (LCIA).12 (ISO. A dynamics simulation model also phased approach that consists of four major components: provides an integrated analysis framework that can include: goal definition and scoping. shortcomings of the existing literature and modeling 2005). EPA. and specifically leverage past and current LCA knowledge base present a challenge in designing scaled work specific to biofuels (ANL 2009. and co-product credits. including the raw material acquisition required to manufacture the product 10. 2009. combination of improved biological productivity and fully integrated production systems can bring the cost The term “life cycle” refers to the major activities in the down to a point where algal biofuels can be competitive course of the product’s life-span. Exhibit 10. While most algal biofuels production. variability in approach to final costs (from per gallon LCA is considered to be a key element of the scope of TE of algal oil to per kg of “raw” biomass) and illustrate modeling and analysis within the context of this roadmap. In resource capture and reuse.In summary. To facilitate the Workshop’s can play a significant role in the national energy economy objectives of identifying needs. 1997. There are indications.g. costs. and numerous others (EPA. would be subject to narrow operating margins. they also present a wide based on other renewable and non-renewable feedstocks. 1997.6 Preliminary System (EPA. These included technical reports prepared environmental impacts and tradeoffs of industrial systems for the U. data base resources. Systems and Techno-Economic Analysis  . and compiling suggested directions for research and development from the participants. 1987)... 2009). with petroleum at approximately $100 per barrel. 1996. and policy issues • Feedbacks and Multiple Sector Interdependencies with links to other models and analyses 104 10. the UC/Davis Lifecycle Emission Model (Delucci.g. • Input resources. a general lack of demonstrated operating parameters The systems modeling and analysis framework for algae and widely varying basic assumptions on a number of biofuels must include the integration of appropriate parameters from algal productivity to capital depreciation LCA methodology and metrics (Curran. and water.

and resource constraints. The model approaches1 currently includes only a limited amount of available data. and has • Employing modular modeling. Interactive graphical user interfaces. data and assumptions from the same information sources discussed earlier in this chapter and in chapter • Determine the current state of technology 9. Workshop effort and is currently a work in progress. • Incorporate appropriate insights and benefit from alliances with industry associations 10. Systems and Techno-Economic Analysis  105 .can facilitate the • Identify critical path elements that offer opportunities use of the model to conduct rapid ‘what if’ analyses and for cost reduction tradeoff studies using different yield scenarios. The preliminary participants specifically suggested that the following modeling uses algae production and resource availability areas be addressed in the modeling and analysis. technical • Identify research areas most in need of support and economic performance for various pathways.7 Potential Directions The Techno-Economic Analysis can accomplish this by: for R&D Effort • Stressing dynamics over detail initially. ISBL and OSBL been developed further since the workshop. e.The development of a system dynamics model for \in techno-economic analysis and the capture of data conducting scenario analysis of algal biofuels production from fielded systems as they emerge over time can scale-up and operations was initiated as part of the roadmap provided needed additional information.11 Scope of Life Cycle Analysis (Adapted from EPA 1993) 10. fuel portfolio along with the ability to do Monte Carlo simulation. The model will eventually include • Identify external factors that will impact cost more detailed systems and processes modules for multiple • Provide plan for entry of algal biofuel into a renewable pathways being pursued for algae biofuels and co-products. continued progress INPUTS OUTPUTS Atmospheric Raw Materials Acquisition Emissions Waterborne Raw Materials Wastes Manufacturing Solid Wastes Use / Reuse / Maintenance Coproducts Energy Recycle / Waste Management Other Releases System Boundary Exhibit 10.g. • Inform and perhaps guide formation and/or varying parameters values within pre-set ranges in order modifications to public policy to describe the uncertainty or robustness of model output. • Establishing interface requirements between To adequately inform research and investment decisions sub-systems for algal biofuel deployment. expanding The model described above was initially prepared in detail later as available and needed outline form for the algae roadmap workshop.

12 Phases of a Life Cycle Analysis (Adapted from ISO. Systems and Techno-Economic Analysis  . the complementary bi-lateral feedback of process performance uncertainties (technical and economic) information and insight between the modeling and analysis. etc. can facilitate Roadmap. Throughout the roadmap workshop and report development When done in a closely coordinated spiral development process. have been identified along various steps in the value chain. 1997) • Leveraging university and industry resources algal biofuels industry. and the experimental and operational investigations These have been noted in this and earlier chapters of the with larger-scale systems in the field. Addressing the uncertainties in a systematic more rapid technological advancement leading toward and integrated modeling and analysis framework will help commercialization. monitoring. Goal Defination and Scope Interpretatation Inventory Impact Analysis Assessment Exhibit 10. guide needed investments and speed the deployment of an 106 10. A critical complementary need is • Maintaining industry standard notation. significant algae-to-biofuels production system and fashion. units. operation. and evaluation of larger-scale algae biofuel production systems and processes that can provide data to validate and improve the modeling. the development.

Bioenergy Research. Biosorption of heavy metals by microalgae in batch and continuous systems. 2004. Critical Reviews in Biotechnology. TP-510-29417. Opportunities for renewable bioenergy using microorganisms. http://www. “Factors Contributing to the Recent Increase in U.A. Thomas-Hall. DOE’s aquatic species program – Biodiesel from algae. Mehta. Report prepared for the Pittsburg Energy Technology Center. (2008).hydrogen.R. ERS Report AR-33. 100(2). (1997).. “Environmental implications of power generation via coal-microalgae cofiring”. S. Mussgnug. Marx..L. and Oswald. 1(1).. Huang. 37-54. Biotechnology Advances.References Aksu. K. Benemann. (2009). Environmental Life Cycle Assessment. E. Bioenergy knowledge discovery framework. Marcel Aillery. P..G. P. AspenTech (2010). “Microalgae Production from Power Plant Flue Gas: Environmental Implications on a Life Cycle Basis”. Available online at: http://www. Environmental Protection Agency (EPA). February 2009. Fertilizer Prices. Systems and Techno-Economic Analysis  107 .P. 1997. Journal of Computational Biology.localenergy. C. A.epa.E. 20. and Chisti. DOE (2009). Scott A. A.H.. (2003). Dunahay. Mark (2004). M. eds. March 2009. G. Cincinnati: Ohio. (2003). pp. (Grant No. Systems and economic analysis of microalgae ponds for conversion of CO2 to biomass. Optimizing algae growth in open-channel raceways. Malcolm. Cincinnati: Ohio. ISBN 0-07-015063-X. and Marca Weinberg (2009). “Ethanol and a Changing Agricultural Landscape”. Springer-Verlag. (2009).gov/ and specifically the DOE Hydrogen Analysis (H2A) Project: http://www. Berlin. Geographic Information Science and Technology Group. The greenhouse gas. McGraw-Hill. 20– 905-922. USDA ERS Report Number 86.L..Life cycle assessment .aspentech.pdf 10. Acien Fernandez. Energy v. Environmental Protection Agency (EPA).S. Mgmt. US-EPA LCA Resources Web Site (latest update July 2009): Available on line at: http://www. “Power Plant Flue Gas as a Source of CO2 for Microalgae Cultivation: Economic Impact of Different Process Options”.F. Available online at: http://www..jsp?query_id=1&page=0&osti_id=493389 Curran (1996). Medina. Environmental management . v. 2002-08”..27.K. J. (1997). Sheehan.anl. 491-515. K. James.hydrogen. http://www. (in press). T. regulated emissions. Y. Kadam. (2002).. J. November 2009.R. (1998).. (1996). C.L. D. ISO 14040. Belarbi.. Y-S Won and N. (2001)..nrel. UCD-ITS-RR-04-45. Recovery of microalgal biomass and metabolites: Process options and economics. Kruse. Wen-yuan (2009). pp. E. (NREL/TP-580-24190). (1993). Oak Ridge National Laboratory (ORNL).osti. (2006). A look back at the U.C.Principles and Framework. Office of Research and Development.. S505-S510. DOE Hydrogen Program. International Standards Delucchi. NREL Report No. http://www. Argonne National Laboratory (2009).Y. B. Molina. DE-FG22- 93PC93204).S. Energy Convers. Kadam.. pp. and energy use in transportation (GREET) model. AspenTech software product portfolio.38. Available online at: http://www. Curran (ed). (2005) Use of algae for removing heavy metal ions from wastewater: Progress and prospects. and Varun. Schenk. including AspenPlus® and Aspen Process Economic Analyzer (formerly Aspen Icarus Process Evaluator®). Posten. Oak Ridge National Laboratory. Life cycle assessment: Principles and practice. Benneman. J. 2002. S. “Conceptual and Methodological Issues in Lifecycle Analyses of Transportation Fuels“. Bioenergy conversion factors. Suppl. Kadam.” In: Wastewater Treatment with Algae.html Getman. K. International Standards Organization.. Second generation biofuels: High-efficiency microalgae for biodiesel production. (1998). B. and Hankamer.transportation. and Roessler. S.pdf Rittmann. Office of Research and Development. (2008)..html Environmental Protection Agency (EPA). Biotechnology and Bioengineering. Tam. (EPA/600/R-92/245). F. USDA. Life Cycle Assessment: Inventory Guidelines and Principles. W. J. and Gaur. (EPA/600/R-06/060).biblio. 25(3). J.H.

108 10.. 1115-1122. 873-885. United Nations Environmental Program. (1987). Systems and Techno-Economic Analysis  . A. B. Life cycle approaches: The road from analysis to practice. UNEP/SETAC Life Cycle Initiative. A. Microalgae production: Technical and economic evaluations. Available online at: http://jp1. D.asp?site=lcinit&page_id=138F5949-6997-4BE6-A553-585E92C22EE4#lcirap Weyer. (2009). Lundquist. 32. Darzins. and Y.. I. R. 1 -10. Bush. Nelson (2009). T. net/sites/lcinit/default. pp.estis.Tapie. M. D. Theoretical Maximum Algal Oil Production. Biotechnology and Bioengineering. n11. American Society of Civil Engineers. & Willson. BioEnergy Research. K. (2005). Feffer. A.. P and Bernard. Journal of Environmental Engineering.. “Algae Grown on Dairy and Municipal Wastewater for Simultaneous Nutrient Removal and Lipid Production for Biofuel Feedstock”. v135. Woertz. United Nations Enivironment Programme (UNEP).

2001.. pre-revenue. given the current state of of a sustainable. including specific studies on the impacts firms. leading public-private partnerships not only for large-scale to rapid industry growth and a stable market. also noted the unique partnership on sharing of knowledge and capabilities for mutual environment in algal biofuels development. The the algal biofuels value chain. Link could.. infrastructure projects.1 shows the potential benefits of for membership and intellectual property models. This partnerships are documented (Audretsch et al.g. 11. algal biofuels industry. Public-Private Partnerships   109 . Gallagher et al. in past efforts are discussed in the context of applicability to the algal biofuels challenge. Brown et al. public- private partnerships have the potential to accelerate Since the 1980s. many focusing on one to a few elements in national laboratories. Indeed. but not necessarily producing transportation envisioned by the Council on Competitiveness (1996): biofuels from cultivated algal biomass at scale. Industry benefits from public-private partnerships from the 1999). 2006). increase the capital efficiency of commercial et al. The type of this industry. By focsuing these partnerships on pre- competitive research or critical problems shared by all Exhibit 11... given benefit could pull together the current expertise and the fact that the algal biofuels industry is still in its facilities.1 The Benefits of Algal Biofuels While benefiting both private and public entities from Public-Private Partnerships shared investment toward mutual objectives. Various models for such partnerships employed of a shared R&D objective. however. This chapter discusses potential models for public-private and government agencies and laboratories in partnerships in general and specifically as related to algal various combinations to pool resources in pursuit biofuels. academia. universities. industry) and public entities (e. the U. Partnerships based participants. and government programs aimed at public-private which can support a quickening pace of innovation. has increasingly invoked commercialization of algal biofuel technology. in turn. collaboration between private entities (e.A 11.. More specifically. as well as reduces the risk of private 2001. analyses of various federal agencies exposure to fundamental science and engineering R&D.11. academia can be either public or private. thereby facilitating growth and development infancy. national laboratories and universities) for development of algal biofuels. many of which may be investor-backed and of DOE programs on the clean energy sector (Brown. 2002. Partnerships are defined as cooperative arrangements engaging companies. investment.g. and industry.. 2002). including characteristics Exhibit 11. but also for research and technology developments of national interest (Stiglitz and Wallsten. the business strategies of many existing partnership most relevant to the advancement of the algal companies are focused on one or more aspects of biofuels industry is perhaps the research partnership algae. realizing benefits in both categories.1 Benefits of algal biofuels public-private partnerships Public Benefits Shared Benefits Private Benefits Energy Security Innovation Quickens Technolgy Commercialization Long-term Clean Fuel Shared Risk & Industry Growth & Market Stability Capital Efficiency Solution Shared Investment Competitive Edge Climate Change Migration Domestic Energy Supply Breakdown Legal & Jobs Creation Sustainable Industry Regulatory Hurdles Clean Energy & Development Science Leadership Growth Stimulates Labor Pool A In this situation.S. Public-Private Partnerships The Workshop participants emphasized the need The algal biofuels industry is evolving with numerous for DOE and other federal agencies to partner with players.

Through their structure.. new partnership. technology scale-up and demonstration. while increasing the opportunity to through public-private partnerships. Basic research and early stage technology with limited funding Networking events such as workshops. particularly in instances in which the team would not At the highest level. which could allow In recent years. as well as a platform to share research. such forums may favor mature • Consortia and trade organizations technology in the final stages of development where the • Team-oriented prizes focus can be on assembly into a working system. of solicitations are system-oriented. • The benefits of partnership outweigh the cost Consortia and trade organizations are membership of collaboration. These events are often 11. and provide publicity • Networking events such as seminars and conferences and marketing toolsets for competitors to attract future • Team-oriented government solicitations investors.2 Components of Successful in a project-level partnership. these organizations provide for opportunity to facilitate deeper collaborations. conferences. including the development of biofuels. Through these events. into a working system to solve a problem. non-profit foundations. However. and potential partners can explore areas of mutual interest. these partnerships can • The partners collaborate on the basis of only be expected to last for the life of the project. global problems. national. and private investors) into a single competition. 110 11. Such solicitations have Public-Private Partnerships the potential to accelerate technology development. as common interest. Often these types develop and license technology with new partners. consortia and trade organizations can also be an effective • The partnership should not openly conflict with the vehicle for researchers looking for a new approach or interest of other groups (Micheau. successful partnerships have have necessarily come together without a government- been identified to include the following attributes: funded solicitation. Prize forums can also leverage reward money There are several forums for creating public-private from a variety of different funding sources (state and partnerships: federal government. or levels beyond the capability of any individual entity. 2008). these environments through collaborations among • The benefits received from the partnership should the membership. be proportional to the value of the contribution. However. By maintaining an active directory. Partnerships can bring together parties that have not organizations that promote a closed circle of partners who worked together before. Public-Private Partnerships   .C Prize forums can pull together developed technology 11.3 Forums for Public. because some awards are based on having solved the problem. labor).B.4 Partnership Challenges in the point at which new researchers can enter and where the Algal Biofuels Industry new thought or methodology is shared with the peers. environment. Implicit in the gained through openness toward new partnerships. industry would retain its collaborative research to tackle complex problems competitive edge. Partnerships are facilitated in than they can individually. Government solicitations are also an option to promote regulatory issues. where no single entity can address the complex research challenge. and demand that the winning team meet or exceed certain success Private Partnerships criteria. concept of collaborating on a common basis is the sharing of pre-competitive research results.g. they are often formed to serve no other purpose. However. scientists and engineers can learn of new research. organizations that may provide an opportunity for • The partners can achieve more through collaboration lasting partnerships. and may compete poorly in this type of award. which could both be a benefit have worked together for many years may not generate (new complementary capability) and a challenge (the the new thinking and fresh approaches that could be understanding of how to work together). prizes have been offered by various for the advancement of technology and know-how to organizations to support solving critical. This may result in the formation of teams of proposers 11. with the ultimate seminars can be important tools for creating a collaborative solution only as good as existing technology will allow.players (e.

technology development. algal growth and harvesting. Algal biofuels will likely face similar issues since natural environmental constraints Different consortia models exist and it is possible to could have a significant impact on algal productivity. or trade organizations—can be an be the workhorse that generates labor for this new industry. These PPP models are scaling barrier and the lack of private capital to conduct mentioned only to serve as examples of the four external such applied research. These include: are needed to engage field experts (e. public-private partnerships the algal biofuels to become viable. this characteristics. nanotechnology. economic. particular algal • Share knowledge of existing algal strains through strain.mitcep.. several in algal biofuels. A compare some of these existing public-private consortia public-private partnership for pre-commercial-scale user models against these four external characteristics. construction personnel. consortia. aviation. develop new algae-based fuels and co-products. etc. conversion process.) or an open access repository and collaborate on the end goals (specific intended use. biofuels.Public-private partnerships—whether they As this industry expands. Public-Private Partnerships   111 . overcoming the lab-field and information technology. Collaborative partnerships. facilities could allow for new technologies to be road- as illustrated in Exhibit 11. collaborate on the development of dynamic and expected contributions of the membership.xprize. 2009). collaborative consortia for technology development are • The generation of basic methods and standards. Does it share investment equitably? field (Willems.5 Modeling Partnerships for • Pre-commercial-scale user facilities that are Technology Development accessible to researchers and developers Of the different forums for public-private partnerships.g. and the most difficult to implement.. marketing • An open environment to share ideas and technology and public relations specialists.g. distributors) across the across the entire algal biofuels value chain. lawyers. particularly between industry and public-policy and regulatory challenges. This type of partnership would greatly accelerate semiconductors. in order to fully realize the potential key technological challenges must be addressed for of the algal biofuels industry.). 11. performance aspects of development of new natural or GMO algal strains the fuel. Therefore. These consortia require • Labor force and intellectual talent to draw upon. etc. the established industry sectors represented by most B http://www. Does it seek to grow the industry? laboratory have traditionally not translated well to the • Shared Investment. Is it structured to develop and commercialize new technology? In the biofuels industry. The exhibit compares tested and scaled before full-scale deployment in the PPP models from five different industry sectors – field. monitoring of growth operations • Collaborate on the development of technologies Considering the following four external characteristics for biomass harvesting and extraction of can help evaluate consortia models: algal components from the biomass • Share knowledge of fuel requirements • Openness.2. events. government solicitations. all of the key attributes for successful partnerships noted earlier. it will need labor to run its are formed or sustained through networking operations. algal oil 11. Academia will competitions. it is useful to conceptualize Technology partnerships could be used for following the various models for public-private consortia in terms objectives: of the five attributes of successful partnerships within the context of particular scenarios (e. and fuel conversion. How inclusive is the membership and utilization to advance fuel conversion to its industry (or segment thereof)? technologies toward marketable products • Technology Commercialization. Because the ecosystem in the algal type of partnership can help incentivize commercial biofuels industry is unique and differs greatly from investment of similar full-scale algal operations. Also. and academia. By reducing scaling C http://www. real estate developers. are needed to ensure that academia develops students capable of addressing the challenges industry faces As discussed in greater detail in this document. requirements. prize and innovate new cost-cutting measures. effective way to address technological. technologies developed in the • Industry Growth. dewatering pathway. to road-test their technologies. • Technology development in algal biology. Doing so may help define the boundary problem(s) for focus by a public-private consortium • Share knowledge of algal cultivation and and bring clarity to the composition. value chain to address technical and non-technical needs.

coordinated royalty-free license to industry roadmap to members focus R&D and spur • PPP can also license IP to on economic growth third parties • R&D conducted at Research • Address game. then adopted development • Now funded facilities amongst members • PPP owns created IP and • Develops solely by industry • Public conferences provides non-exclusive. Public-Private Partnerships   . discuss and debate the merits of each. and determine how to best adapt / implement these attributes. • Government Government Centers or partner facilities changing. However.models presented. this industry may find that no specific model will meet all of its needs.2 Comparison of Some Public-Private-Partnership (PPP) Models TECHNOLOGY SHARED INDUSTRY FOCUS ENTITY TYPE OPENNESS OF PPP INDUSTRY GROWTH COMMERCIALIZATION INVESTMENT • Collaborate on pre-competitive R&D • Designed selected by membership to increase competitiveness of • Transfer of technology by existing US firms in publication or member- marketplace • Open membership website data transfer • Provides • Cost-shared Non-Profit • Represent • Technology further commercialization by government Corporation industry interests developed to manufacturing network that and industry at Semiconductors (501c6) with • Forums inspiring solutions with external drives economic start-up physical cross collaboration partners. it could be helpful to consider these attributes and models. for an algal biofuels public-private consortium to meet its specific mission successfully. Exhibit 11. high-risk Agency • Partners with barriers through funding Research industry and • Industry partners to targeted R&D • State funding Centers universities commercialize developed IP • Industry growth • Financial and Biofuels (not a • Public conferences • IP licensed to interested supported through other resources separate legal and workshops parties with an evaluation education from national entity with of commercialization labs • Opportunities for potential • Limited industry employees) collaboration involvement to • Transfer of technology by address issues on publication industry growth 112 11.

non-exclusively general aviation funding for over 8 with NASA • Consortium of • Commercialization afforded market. • Industry growth funding for over 5 • Research conducted by Corporation • Technical through education years national labs and member Nanotechnology (501c6) with workshops. facilities business. businesses of years employees exclusively to each other • Significant industry royalty-free involvement • Transfer of technology by provides market publication. and transfer focus and of knowledge through commercialization member-only database network • Donor-driven • Research conducted by model • Industry growth university • Industry and supported through University • Software is open source • State funding university partners education of California licensed over 4 years Information • Numerous forums • Limited industry Institute • Other IP is either licensed • University funds Technology Research inspiring cross involvement (not a separate non-exclusively. • Technology • Founding universities physical technical. and regulatory designed to reduce bottlenecks operational costs Industry • IP licensed exclusively or and increase • Government consortium • Open membership. academic as members agree to cross large established • Industry match separate legal and government license background and firms and small for same number entity with partners newly developed IP non. standardization operational. including years Aviation & FAA (not a industry. Public-Private Partnerships   113 . • Industry gifts collaboration through Advisory legal entity free basis or exclusively. • No federal amongst committee to with employees royalty basis as needed to funding researchers from address issues on achieve the widest possible universities and industry growth dissemination. companies 11. and members commitment for paid-up license to industry social issues over 3 years members inspire cross collaboration amongst members • Collaborative research amongst consortium • Technology members designed development and to reduce technical. academic • Pre-competitive research and national lab • Government selected by all members Non-Profit partners. royalty. TECHNOLOGY SHARED INDUSTRY FOCUS ENTITY TYPE OPENNESS OF PPP INDUSTRY GROWTH COMMERCIALIZATION INVESTMENT • Open membership • Consortium of industry. only licensed to member financial • PPP provides non-exclusive.

A. J. Willem. W.N. 325(5941). Market failures and barriers as a basis for clean energy policies. Research Policy. Energy-technology innovation. (1996).A. (2009).S.V. Public-Private Partnerships   . Annual Review of Environment and Resources. 707-708. Brown.. M. Energy Policy. Gallagher.A. D. An analysis of public initiatives to promote strategic research partnerships. (2001). Public/private technology partnerships: evaluating SBIR-supported research. J. 31. Washington: U. 1197-1207. Paton. A. D. J.A. Link.G. Short. S.P. Science. and Koomey J. A. 31. 29(14). Public-private policy partnerships. Endless frontiers. (2000).. Link..D. and Sagar. D. M.D. K.. P. (2002).. (2002). R&D Policy for Competitiveness. Scenarios for a clean energy future.T. (2006).J.. In Rosenau. and Scott. 29(14).S.References Audretsch. 145-158. and Wallsten. 114 11. Brown. (2001). M. 1179-1196. Council on Competitiveness. Stiglitz. 31(1). The biofuels landscape through the lens of industrial chemistry.S.. and Siegel. The MIT Press.N. Research Policy. Energy Policy.. Holdren.. limited resources.B. 1459-1466. P.E. Public-private technology partnerships: Promises and pitflass. 193-237. (ed). Levine.

S. Inc. Department of Energy David Bayless Ohio University Jacques Beaudry-Losique U.Appendices Appendix 1 : Workshop Participants Walter Adey Smithsonian Institution John Aikens Lybradyn Mark Allen A2be Carbon Capture Gennady Ananyev Princeton Alex Aravanis Sapphire Energy Ed Archuleta El Paso Water Utilities Paul Argyropoulos Environmental Protection Agency Ricardo Arjona Abengoa Bioenergy Virginia Armbrust University of Washington Bob Avant Texas A&M Tom Avedisian U.S. Andrew Braff Wilson Sonsini Goodrich & Rosati Jim Brainard National Renewable Energy Laboratory Jerry Brand University of Texas at Austin Tom Brennan Arch Ventures Michael Briggs University of New Hampshire Jim Bristow Joint Genome Institute Kevin Brown - Malcolm Brown University of Texas at Austin Nathan Brown Federal Aviation Administration (Office of Environment And Energy) Kent Bryan City of Celina 115 Appendices   . Diplomacy Isaac Berzin And Strategy Catalino Blanche U.S. Department of Energy Biomass Program Leslie Bebout NASA Ames Research Center Lucie Bednarova General Motors Amha Belay Earthrise Nutritionals John Benemann Benemann Associates Linda Benjamin Food and Drug Administration Christoph Benning Michigan State Interdisciplinary Center Herzliya. Department of Agriculture Doug Borger Lockheed Martin John Bouterse Envo Smart Sara Boyd BCS. Lauder School of Government.

Department of Agriculture Paul Falkowski Rutgers University John Ferrell U. Department of Energy. Department of Agriculture Heriberto Cerutti University of Nebraska . Department of Energy Biomass Program Michel Delafontaine Hawaii Biocrude Gregg Deluga General Electric Global Research Harrison Dillon Solazyme Dan Drell Office of Science Biological and Environmental Research Donn Dresselhuys Old Dominion University Bruce Eldridge University of Texas at Austin Tj Evens U. Das University of Georgia John Datko Air Force Office of Scientific Research Chris Davies Independence Bioproducts Brian Davison Oak Ridge National Laboratory Mark Decot U.S.S.S. Todd French Mississippi State Christian Fritsen Desert Research Institute Matthew Frome Solozyme Maria Ghirardi National Renewable Energy Laboratory Appendices   116 .William Buchan NASA Ames Research Center Jay Burns Bionavitas Arthur Butcher CPS Energy Thomas Byrne Byrne & Co Martin Carrera BP Matt Caspari Aurora Biofuels Chris Cassidy U.Lincoln Alice Chen Environmental Protection Agency Parag Chitnis National Science Foundation Daniel Cicero National Energy Technology Laboratory Mike Cleary National Renewable Energy Laboratory John Coleman Algenol Jim Cook Univenture Doris Cooksey CPS Energy Keith Cooksey Montana State University Michael Cooney University of Hawaii – Manoa David Daggett Boeing Al Darzins National Renewable Energy Laboratory K. Biomass Program Daniel Fishman BCS.C. Inc.S.

Department of Agriculture Tom Gross If.S. Department of Energy. Department of Agriculture Mark Hildebrand Scripps Institute Masahito Hiratake Mitsubishi International Kent Hoekman Desert Research Institute Neil Hoffman U.S. Department of Energy. Biomass Program Michael Graves University of Massachusetts Bailey Green Lawrence Berkeley National Laboratory Eli Greenbaum Oak Ridge National Laboratory Nathanael Greene Natural Resources Defense Council Michael Gregoire U.Warren Gillette Federal Aviation Administration (Office of Environment And Energy) Jim Gillingham Valero Dan Ginosar Lawrence Livermore National Laboratory Robert Glass Lawrence Livermore National Laboratory Steven Gluck Dow Chemical Steve Gorin National Renewable Energy Laboratory Alison Goss Eng U. LLC Arthur Grossman Stanford Guritno “Guri” Roesijadi Pacific Northwest National Laboratory Molly Hames U. Biomass Program Suzanne Goss Jacksonville Electric Authority Paul Grabowski U. Department of Energy.S. Department of Agriculture Ed Hogan Natural Resources Canada John Hogan NASA Ames Research Center John Holladay Pacific Northwest National Laboratory 117 Appendices   .S.S.S. Department of Energy Biomass Program Laurel Harmon UOP Deanna Hatch Organofuels Incorporated Pat Hatcher Old Dominion University David Hazlebeck General Atomics Bob Hebner University of Texas at Austin Grant Heffelfinger Sandia National Laboratories Erik Helm Environmental Protection Agency William Henley Oklahoma State Doug Henston Solix Biofuels Jurg Heuberger County of Imperial Kevin Hicks U. Golden Field Office Anders Handlos Grauslund Novozymes Zia Haq U.S.

Inc Qiang Hu Arizona State University Neil Huber Army Michael Huesemann Pacific Northwest National Laboratory Peter Hug Recombinant Innovation.S. Hunter-Cevera University of Maryland Biotechnology Institute Mark Huntley University of Hawaii Stephen J. Rachel Lattimore Arent Fox LLP Edward Laws University of Hawaii at Monoa Gordon Leblanc Petrosun Joo-Youp Lee University of Cincinnati Paul Lefebvre University of Minnesota James Lester Stratus Consulting Jack Lewnard Gas Technology Institute Casey Lippmeier Martek Kent Lohman Midwest Research Institute Tryg Lundquist California Polytechnic State University Appendices   118 .C.Andy Horne Natural Resources Development Jeff Houff Hydrogeologic. LLC J. Department of Energy Biomass Program Geetha Kothandaraman ConocoPhillips Walt Kozumbo Air Force Office of Scientific Research Heidi Kuehnle Kuehnle Agrosystems Peter Lammers New Mexico State University Cynthia Lane American Water Works Association Todd Lane Sandia National Laboratories Mario Larach Kai Bioenergy Corp. Miller Chevron Chevron Gil Jackson United States Agency for International Development Jacob Jacobson Idaho National Laboratory Robert Jagusch Minnesota Muncipal Utilitees Association Eric Jarvis National Renewable Energy Laboratory Jeff Jeter Honda Libby Jewett National Oceanic and Atmospheric Administration James “Jim” Sears A2be Carbon Capture Michael Johnson Massachusetts Institute of Technology Glen Kertz Valcent Alex Kinnier Khosla Ventures Douglas Kirkpatrick Defense Advanced Research Projects Agency Melissa Klembara U.

Kenneth Moss Environmental Protection Agency Chuck Mueller Sandia National Laboratories Jeff Muhs Utah State University Walter Mulbry U. Mayfield Scripps Research Institute Mike Pacheco National Renewable Energy Laboratory Paul Panesgrau National Renewable Energy Laboratory Frank Parker Organofuels Incorporated Helen Parker Algaelink.John Magnuson Pacific Northwest National Laboratory Len Malczynski Sandia National Laboratories Mike Manella ADM Russell Martin Blue Source Anthony Martino Sandia National Laboratories Margaret McCormick Targeted Growth Patrick McGinn National Research Council John McGowen Biodesign Institute at Arizona State University Diane McKnight University of Colorado-Boulder Steve Meller Procter & Gamble Omar Mendoza United States Air Force Walter Messier Evolutionary Genomics Blaine Metting Pacific Northwest National Laboratory Anelia Milbrandt National Renewable Energy Laboratory Greg Mitchell Scripps Institution of Oceanography John Mizroch U. Department of Energy Lissa Morgenthaler-Jones Livefuels Jerry Morris Innovation/Biodomain .Shell Global Solutions Inc.V. Roberta Parry Environmental Protection Agency 119 Appendices   .S.S. N. Department of Agriculture Alyson Myers Kegotank Biofuels Nick Nagle National Renewable Energy Laboratory David Nobles University of Texas at Austin Charley O’kelly Blue Marble Energy Lou Ogaard CEHMM Kelly Ogilvie Blue Marble Energy John Ohlrogge Michigan State University Jose Olivares Los Alamos National Laboratory Phil Ovitt Senator Brownback Office George Oyler Johns Hopkins University Stephen P.

Biomass Program Schild Philippe European Commission Philip Pienkos National Renewable Energy Laboratory Maria Piera Abengoa Bioenergy John Pierce Wilson Sonsini Goodrich & Rosati Daniel Pike AF Research Lab Jennifer Pinkerton LA Department of Water & Power Bradley Plantz University of Nebraska-Lincoln John Plaza John Plaza Robert Polak Phycal Juergen Polle Brooklyn College Matt Posewitz Colorado School of Mines Steve Potts EPA Office of Water Peter Pryfogle Idaho National Laboratory Steve Przesmitski National Renewable Energy Laboratory Theras Pugh American Public Power Association Ron Putt Auburn University Nigel Quinn Lawrence Berkeley National Laboratory Kenneth Reardon Colorado State University Hal Reed American Water Valerie Reed U.S.S. Biomass Program Ron Reinsfelder Shell Global Solutions Bruce Resnick Cargill Curt Rich Van Ness Feldman Mark Riedy Andrews Kurth Kathy Roach MurphyTate. LLC John Robbins Exxonmobil Paul Roessler Synthetic Genomics Roger Ruan University of Minnesota Jim Ryan MOR Supercritical. Department of Energy. Department of Energy.Ron Pate Sandia National Laboratories Leslie Pezzullo U. LLC Gerson Santos-Leon Abengoa Bioenergy Richard Sayre Donald Danforth Plant Science Center Philippe Schild European Commission Gerald Seeley Old Dominion University/Company Mark Segal Environmental Protection Agency Mike Seibert National Renewable Energy Laboratory Joel Serface Kleiner Perkins Appendices   120 .

V. LLC Tanja Shonkwiler Jamestown Ny (Outside Counsel) Blake Simmons Sandia National Laboratories Knut Simonsen DTE Energy Resources.S. Biomass Program John Spear Colorado School of Mines Karl Stahlkopf Hawaiian Electric Company. Department of Energy. Cai Steger Natural Resources Defense Council Dan Stepan Energy & Environmental Research Center Jake Stewart Organic Fuels Mark Stone Desert Research Institute Belinda Sturm Kansas University Transportation Research Institute Sidhu Sukh University of Dayton Xiaolei Sun Arizona Public Service Sam Tagore U.S. Inc.Pamela Serino Defense Energy Support Center Chris Shaddix Sandia National Laboratories Alex Shearer SRI International John Sheehan Sheehanboyce. Biomass Program Evonne Tang National Academy of Sciences Jeff Tester Massachusetts Institute of Technology Robert Thomas South Australia R&D Institute Jonathan Trent NASA Ames Research Center Pat Unkefer Los Alamos James Van Etten University of Nebraska Peter Van Den Dorpel Algaelink. Inc. Department of Energy.Lincoln Alan Weimer University of Colorado at Boulder Zhiyou Wen Virginia Tech Norman Whitton Sunrise Ridge Algae 121 Appendices   . Evan Smith The Verno Group Seth Snyder Argonne National Laboratory Milton Sommerfeld Arizona State University Glenn Sonntag U. N. Jean Vandergheynst UC Davis Tom Verry National Biodiesel Board Sanjay Wagle Vantagepoint Venture Kevin Wanttaja Salt River Project Michael Webber The University of Texas at Austin Alan Weber Marc-IV Consulting Donald Weeks University of Nebraska .

LLC. Appendices   122 . Inc.Elizabeth Willet Mars Symbioscience Bryan Willson Colorado State University Byard Wood Utah State University Christopher Wood Platt River Power Authority David Woodbury Biofuel Producers of America (Bpa Paul Woods Algenol Ben Wu Livefuels Don Wyma National Intelligence Officer For Science And Technology Joyce Yang U. Inc. Maria Ghirardi National Renewable Energy Laboratory Stephen Gorin National Renewable Energy Laboratory Tom Gross IF.S. Molly Hames U. Department of Energy Golden Field Office Grant Heffelfinger Sandia National Laboratories Mark Hildebrand Scripps Institution of Oceanography John Hogan NASA Ames Research Center Qiang Hu Arizona State University Michael Huesemann Pacific Northwest National Laboratory Eric Jarvis National Renewable Energy Laboratory Geoff Klise Sandia National Laboratories Alina Kulikowski-Tan Carbon Capture Corp Pete Lammers New Mexico State University Jared Largen BCS.S. Department of Energy Biomass Program Chris Yeager Savannah River National Laboratory Ross Youngs Univenture Tim Zenk Sapphire Energy Zhenqi Zhu Stevens Institute of Technology Yonathan Zohar University of Maryland Biotechnology Institute Appendix 2: Contributors Jim Brainard National Renewable Energy Laboratory Tom Brennan Arch Ventures Bill Buchan NASA Ames Research Center Mike Cleary National Renewable Energy Laboratory Michael Cooney University of Hawaii – Manoa Al Darzins National Renewable Energy Laboratory Daniel Fishman BCS.

S. Department of Energy. Department of Energy. Biomass Program Chuck Mueller Sandia National Laboratories Nick Nagle National Renewable Energy Laboratory Michael Pacheco National Renewable Energy Laboratory Ron Pate Sandia National Laboratory Leslie Pezzullo U. Inc. Len Malcynski Sandia National Laboratories Tony Martino National Renewable Energy Laboratory Steve Mayfield Scripps Institution of Oceanography Anelia Milbrandt National Renewable Energy Laboratory Joanne Morello U.S.Yantao Li Arizona State University Tryg Lundquist California Polytechnic State University Rajita Majumdar BCS. LLC. Biomass Program Philip Pienkos National Renewable Energy Laboratory Juergen Polle Brooklyn College Matthew Posewitz Colorado School of Mines Ron Putt Auburn University Kathy Roach MurphyTate. Biomass Program 123 Appendices   .S. Guri Roesijadi Pacific Northwest National Laboratory Mike Seibert National Renewable Energy Laboratory Joel Serface Kleiner Perkins Chris Shaddix Sandia National Laboratories Blake Simmons Sandia National Laboratories Amy Sun Sandia National Laboratories Joyce Yang U. Department of Energy.

Inc. Rittmann Arizona State University G. Solazyme (Chief of Genetics) Douglas Haughey not listed Joel E. Kostka Florida State University David Lewis The University of Adelaide Margaret McCormick Targeted Growth Lissa Morgenthaler-Jones LiveFuels. Bob Avant et al Texas AM Halil Berberoglu University of Texas Shulin Chen et al Washington University Barry Cohen National Algae Association Anju Dahiya 1) The University of Vermont . Algal Farms Inc. Dr. Paul Woods Algenol Xiaohan Yang Oak Ridge National Laboratory Tim Zenk Sapphire Energy Appendices   124 . Biodesign Institute. Inc. LLC Carnegie Institution for Science (Staff member).Appendix 3: Respondents to Request for Information on the Draft Roadmap Daniel Angell TPA Inc. Biotechnology Institute David L. Bruce E. Whitton. Dr. School of Life Sciences Marc von Keitz and Steve Heilmann University of Minnesota. Valentin Soloiu Georgia Southern University Lance Stokes ECI Environmental Compliance Jefferson Tester Cornell University Willem Vermaas Arizona State University. Shenderov BioLogical Technologies. Roesijadi Pacific Northwest National Laboratory Mary Rosenthal Algal Biomass Organization Richard Sayre and Ursula Goodenough Donald Danforth Plant Science Center and Washington University Gerard Seeley Jr. Mark Segal Environmental Protection Agency Office of Pollution Prevention and Toxics Alexander D. Wenbert The Eden Tube Project Norman M. Stanford University Arthur Grossman (Adjunct Professor). 2) General Systems Research LLC Matthew Frome Solazyme Michael R. Inc Matt Peak Prize Capital María Piera Alberola Abengoa Matthew Posewitz Colorado School of Mines Bruce Resnick Cargill Center for Environmental Biotechnology. Robert S Weber Sunrise Ridge Algae. Gale PetroAlgae.


eere. DOE/EE-0332 .