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Mamta Awasthi and Rajiv Kumar Singh, 2011

INT J CURR SCI 2011, 1: 14-23

Development of algae for the production of bioethanol, biomethane, biohydrogen and biodiesel
Mamta Awasthia* and Rajiv Kumar Singhb

Centre for Energy and Environment, National Institute of Technology, Hamirpur
Himachal Pradesh-177 005, India


Department of Botany, Rajiv Gandhi University, Itanagar, Arunachal Pradesh-791 112, India
*Corresponding author: E-mail:

Microalgal biofuels are a viable alternative for clean, economical and sustainable sources of energy. The research and development of
micro-algae needs a massive boost to ease the technical difficulties to overcome the large cost advantage of other biofuel feed stocks. It is a
large source of biomass on non-arable lands and capture of CO2. Lipids produced from algae contain saturated and polar lipids, which are
suitable for use as a fuel feedstock and it exceeds the best producing oil crops. It has been found that representatives of green and diatom
microalgae are the most promising producers of TAGs, the highest content of which (40–60%) has been found in diatom microalgae. This
paper discusses current knowledge on different techniques for microalgal biomass production, harvesting and lipid extraction methods. The
paper also discusses biodiesel production via transesterification of the lipids and other biofuels like biomethane, biodiesel, bioethanol and
biohydrogen depending on the species as well as the biorefinery approach. Besides lipid extraction, the byproducts of processed algae can
be utilized for many other purposes.
Key words: biofuel, bioethanol, biodiesel, bioenergy, microalgae cultivation, harvesting, lipids
Received: 02nd September; Revised: 25th September; Accepted: 19th October; © IJCS New Liberty Group 2011
atmosphere from many stationary emitters by feeding the

carbon-rich flue gas to the algae (Ackman et al., 1968; Adey and

The concept of using micro algae for energy production is not

Loveland, 2007). Algae are able to fix approximately 1.8 kg of

new. Algae require much less water than the traditional cereals,

CO2 for every 1 kg of algae biomass produced (Antoni et al.,

produce more biomass (Table 1) as well as it can be grown in

2007). Approximately 40 ha of algae ponds are required to fix

salt water or in sewage water. Microalgae do not belong to the

the carbon emitted from one MW of power generated from a

category of traditional raw food materials. One of the possible

coal plant (Becker, 1994). Treated wastewater rich in nitrogen

ways in which the cost of biofuel obtained from microalgae can

and phosphorus can be utilized by algae, thereby providing the

be reduced is by adopting biorefinery approach and using wastes

co-benefit of producing biofuels and removing nitrogen and

from other production processes for cultivating them. With the

phosphorus (Bilanovic et al., 1988; Benemann and Oswald,

latest technologies it has become possible to cultivate the

1996; Bigogno et al., 2002).

biomass of algae on a large scale all year round, not only under

Algae cultivation

the conditions of tropical and subtropical climate, but also in

The necessary technology for developing profitable algae-based

zones with moderate climate, even at negative temperatures of

fuel is still in various states of development and the final

outdoor air in winter. There is potential to effectively reduce the

configuration is yet to be determined and demonstrated at the

amount of carbon dioxide and nitrogen oxides released into the

industrial scale. The high capital cost associated with

or by Crops Annual yield (tons/ha) Sugar cane 54-125 Sweet sorghum 35-70 Soybean 1. Open pond systems are shallow ponds in which algae 2008).0 Sweet potato 10-40 Trees 20-50 Microalgae harvesting a batch at a time (like polyethylene bag cultivation). However. flocculation are cultivated. most harvesting methods still involve motion by paddle wheels or rotating structures. properties of the cellular level. Table 1. 1997). such as a high energy cost. two types can solid support for removing nutrients from animal wastewater be used: turbidostat and chemostat culture. or are harvesting of algal cells by flocculation is more convenient than made up of an undefined mixture of strains (Borowitzka. High-density algal to inadequate illuminance. Csordas and Wang. Algal flocculant toxicity. 1993. The flocculating reactions of an algal biomass which allows control over illumination. temperature. semi-batch. produced in closely-controlled laboratory methods to less Harvesting Algae predictable methods in outdoor tanks. an inclined triangular tubular photo bioreactor. In a continuous system. Open pond systems Conventional processes used to harvest micro-algae include concentration through centrifugation (Briens et al. Bosma et al. continuous stirred tank reactors (CSTR). foam fractionation (Brown et al. or aluminum sulfate followed by certain cationic polyelectrolyte ponds covered with green house or usually a photobioreactor (Conover.. The cultures can be defined (one or more selected strains). contamination with predators and other competing algae surface.. centrifugation or gravity filtration. The water is typically kept in et al. 1993). bags. (ATS) can be used in which benthic algae grow on the surface of and continuous systems. Source: Richmond (1986) A photo bioreactor is equipment that is used to harvest algae. Boichenko and Hoffmann. 2001). 1988. because it allows large Chaumont. concentrations of the flocculants and divalent cations. Photo bioreactors can be set up to be continually harvested (the majority of the larger cultivation systems). Some photo bioreactors types include: tubular photo bioreactors. flat-plated photo bioreactors.Mamta Awasthi and Rajiv Kumar Singh.. helical coils made of plastic tubing placed across a column-like structure. etc. 2007) and ultrasonic separation (Chynoweth sewage/water treatment plants. Algae can be (Bridgwater et al.1-4. 2003). and some mixing economic or technical drawbacks. Heterotrophic cultivation of micro cultures can be concentrated by either chemical flocculation or algae for lipids production does not involve CO2 mitigation and centrifugation. 2011 producing microalgae in closed culture systems is the main wastewater treatment programme along with production of algal challenge for commercialization of such systems. Cultivation can be conducted in batch. Canakci and Van Gerpen. membrane from nearby land areas or by channelling the water from filtration (Chisti. or non-feasibility of scaling-up. 1996. 1994). . rectangular tanks.. 1975). can be accomplished by appropriately designed guides. square tubular reactors consist of plastic tubing arranged in a series of squares. 1999. Researchers also took the path of creating ionic strength of the culture solution and other factors (Craggs heterotrophic strains of algae from obligate photoautotrophs due et al. can be used. A variety of chemicals have Indoor culture / Closed culture been tested as flocculants and the most effective was found to be Vessels such as tubes.. 2004). 800-1600 Two methods of cultivation are used for realizing the Algal Turf Scrubber (ATS) biosynthesis abilities of natural and modified strains of In addition to the open ponds or photo bioreactors. Nutrients can be provided through runoff water (Chaumont 1993. carboys. Biomass yields of microalgae versus conventional crops. quantities of culture to be treated. an autotrophic microalgae: in photobioreactors (PBRs) and in open attached algal culture system such as an algal turf scrubber cultivators.. flasks. nutrient are particularly sensitive to the pH. biofuel.

A more efficient method would make use of the low grade waste heat from the power plant to dry the algae contained in a vessel. and other. Lipid content also varies with season as tested in Chlorella vulgaris. (Gordillo et al. This implies a great advantage of the Crop Oil yield (gal/acre-yr) attached algal culture system in terms of ease of biomass Corn 18 harvesting (Das and Veziroglu.. (figure 2 Unpublished data). 1998. The algae paste obtained from filtration/centrifugation contains as much as ca.Mamta Awasthi and Rajiv Kumar Singh. with 60% of total cell dry matter) depending on the species and fine bubbles supplied by an air compressor. However. 50% water content is necessary to produce a solid material that can be easily handled. Microalgal strains with high sodium. chromium. Microalgae as biological sources of lipids and hydrocarbons Ultrasound based methods of algae harvesting are currently under development. 2000). 1. treated analogous to vegetable oil. Alum is a common growth conditions (Dote et al. The biomass harvested from the attached culture system is paste-like pulpy slurry having a water content to that of the cell pellet centrifuged from the including some Chlorella species (FAO. .200 oil) Microalgae (50% 10. Johnson and Wen. Parietochloris incisa (Haesman et al. 2005) Nannochloris sp. 2000) and Botryococcus braunii (Harris. Dewatering/drying process reduces the water content of the algae prior to oil extraction process. In comparison with SVO. A few micro algal species. 1942.000 oil) Energy production from algae Lipids can be used as a liquid fuel in adapted engines Algal lipids (so-called pseudo vegetable oil or PVO) as Straight Vegetable Oil (SVO). Lipids are one of the main components of micro algae (2- flocculation.. it requires a considerable area of land. It uses alum to flocculate an algae/air mixture. is used commercially in grains and timber drying. 2001) Dunaliella species (Gaffron and Rubin. 1997. 2010). or iron and a univalent metal such as potassium or metabolites and sources of energy. Gerpen. oil or lipid content are of great interest in the search for a Algal drying sustainable feedstock for biodiesel. algal are much more heterogeneous than the seed-oil lipid.. have been reported to have the capacity of accumulating large quantities of lipids in cells under favorable conditions. (2009) suspension culture system. Therefore oil is unsaturated to a larger degree making it less appropriate makes it difficult to characterize algae-derived fuel products and for direct combustion in sensitive engines (Table 3). a popular and inexpensive method. Dissolved Air Flotation (DAF) separates algae oil per unit of land in a year compared to terrestrial crops (Table from its culture using features of both froth flotation and 2). 1994). additional methods are currently Algae are far more oil-rich and offer a higher yield of being developed. Energy production pathway can be expressed diagrammatically as shown in figure 1. Fukuda et al. Cotton 35 Soybean 48 Canola 127 Jatropha 202 Oil palm 635 Fig. Ghirardi et al. Euglena gracilis and Chlamydomonas sp. 2011 Froth flotation is a method of separating algae from the medium by adjusting pH and bubbling air through a column to create a froth of algae that accumulates above liquid level. however. Table 2. Solar drying. Microalgae contain lipids name for several trivalent sulfates of metal such as aluminum. Oil output of different biofuel feed stocks Source: Khan et al.. for example AlK(SO4)2. Drying algae to ca. 1989. 90% water content. storage products.. Energy production pathway Microalgae (15% 1. 2001). and fatty acids as membrane components.

1984.. on both lipid content and lipid type. oil can also be . The effects of growth conditions.. At the lower temperature ranges. an increase in fatty acid unsaturation has been observed. Benzene and ether have been used. Li et al.Mamta Awasthi and Rajiv Kumar Singh. 1998. Figure 3. 2000). 2009).. and media composition) affect the quantity and types of lipids as well as fatty acids compositions (Klass. Lipid Content (% of DW) Khan et al. The effect of nitrogen on the lipid fraction bonds Palmitic acid 16:0 12–1 Palmitoleic acid 16:1 55–7 Stearic acid 18:0 1–2 Oleic acid 18:1 58–60 Linoleic acid 18:2 4–20 Linolenic acid 18:3 14–30 and on cell growth of the strain Nannochloris cultured under saline conditions is summarized in Table 4. growth stage of algal cultures and the taxonomic position of algae. Changes in lipid content in different algae containing shorter-chain fatty acids (16 to 18 carbon length) which are ideally suited for biodiesel production (Kadam. nitrogen limitation as in case of Neochloris oleoabundans (Maeda et al. Liu et al... Table 3.. The While more efficient processes are emerging as shown in figure average lipid content of algal cells varies between 1% and 70 % 3. 2008). a simple process is to use a press to extract a large percentage but can reach 90% of dry weight under certain conditions (70-75%) of the oils out of algae. However the triglycerides produced by P.and di-galactosyldiglycerides. Light enhances the formation of polyunsaturated C16 and C18 fatty acids as well as mono. 18 and most of its fatty acids are saturated fatty acid in the range of 16-20 carbons (Gordillo et al. changes such and as temperature and light intensity.. Neochloris oleoabundans is a freshwater Chlorella vulgaris Chlamydomonas sp Euglena gracilis 16 14 12 10 making them less desirable for biodiesel production.incisa is rich of polyunsaturated fatty acid (Haesman et al. stage seasonal of growth..g. Under unfavorable conditions mixed with cyclo-hexane to extract the remaining oil content. 2. chemicals. The remaining pulp can be (Livansky. of growth. 1998. 2005. Energy conversion processes from microalgae Factors affecting oil production in algae Taxonomic environmental June Time Period et al. 20 species that produces up to 80% triglycerides of its total lipids. (e.. 2008). Knuckey et al.g. 2000). (2009)] factors groupings. ideal for biodiesel production. are discussed (Piorreck and Pohl. 1995). sphingolipids and phosphoglycerides in Euglena gracilis and Chlorella vulgaris (Melis et al. 2011 some members of these algal group like Chlorella is known for Fig. Lee et al. 2002.. Lipid accumulation in algae typically occurs during periods of environmental stress. 1998). 2006) as shown in figure 2. Botryococcus enters a stage in which its Chemical methods: Algal oil can also be extracted using unsaponifiable lipid content increases to a level of 90%. Fatty acid composition of microalgal oil [Source: Meng 8 April May Fatty acid July August September The total amount and relative proportion of fatty acids Chain Oil length:no. composition of double (w/total lipid) can be affected by nutritional and environmental factors e. Extraction of algal oil including growth under nutrient-deficient conditions.

An anaerobic digester contains synergistic microbial populations to convert a variety of organic substrates to methane and carbon dioxide.39 42.6 algae. Biodiesel from algal oil Raw microalgal oil is high in viscosity. protein. Biochemical conversion this extraction process is estimated to be much greater than hexane extraction.Ultrasonic extraction can greatly industry and is relatively inexpensive. Alkali-catalyzed transesterification is about 4000 times faster than the acid catalyzed reaction and hence most frequently used commercially (Miao and Wu. Thus algal organic compounds (lipid. this liquified fluid then acts as the solvent in extracting the oil. After the oil has been extracted using an expeller. Supercritical fluid extraction: In supercritical fluid / CO2 extraction. when these bubbles collapse near the cell walls. 2011 separated by hexane extraction. extraction and supercritical fluid extraction method. The oil dissolves in the cyclohexane. CO2 is liquefied under pressure and heated to the point that it has the properties of both a liquid and a gas. These two stages (cold of algae thereby causing the floating up of oil. with an organic solvent such as hexane or petroleum ether. Three chemical methods are hexane solvent method.Mamta Awasthi and Rajiv Kumar Singh. 2009).5 32. Cell growth Internal lipid (mM) (g/L) content (g/L) 0.. or percolation.Osmotic Shock is a sudden reduction in osmotic pressure.6 33. such as oil. The use of acid catalyst has found to be useful but the reaction rates for converting triglycerides to methyl esters are too slow (Meng et al. usually methanol. it creates shock Hexane solvent method: Hexane solvent extraction can be used waves and liquid jets that cause those cells walls to break and in isolation or it can be used along with the oil press/expeller release their contents into the solvent.9 + feeding 2. 2005). which is widely used in the food Ultrasonic-assisted extraction . The method press and hexane solvent) together may derive more than 95% of developed on the basis of acid catalysis allows the processes of the total oil present in the algae. accelerate extraction processes. Transesterfication is the process of converting raw microalgal lipid (triacyleglycerols / free fatty Soxhlet extraction: Soxhlet extraction is an extraction method that uses chemical solvents.9 2. and suspension in an alternating electromagnetic field with varying the pulp is filtered out from the solution. non-toxic and biodegradable biodiesel. However. 2006). Products of the reaction are fatty acid methyl esters (FAME) and glycerol. Osmotic shock is sometimes used to release cellular components. or carbohydrate) .9 0. the cost of acids) to give renewable.Enzymatic extraction uses enzymes to degrade the cell walls with water acting as the solvent. 2009). Other lesser-known extraction methods Enzymatic extraction . Oils from the algae are extracted through repeated washing. Acid catalysis is suitable for transesterification of oils containing high levels of free fatty acids (Metzger and Largeau. Transesterification is a reaction of the parent oil with a short chain alcohol.9 9. The need to subject method. causing pressure in a solution to rupture.. The oil and the pH value by adding CO2 resulting in the cell wall destruction cyclohexane are separated by distillation. in the presence of a catalyst. extracting lipids from microalgae and obtaining biodiesel fuel to Table 4. under reflux in special glassware. Effect of nitrogen concentration (as KNO3) on lipid be combined in a single stage (Meng et al. Osmotic shock .4 9. thus requiring conversion to lower molecular weight constituents in the form of fatty acid alkyl esters. Using an ultrasonic reactor. Biomethane production: The production of biogas from biomass is gaining increasing importance worldwide. soxhlet ultrasonic waves are used to create bubbles in a solvent material. this makes fractionation of the oil much easier. The culture can be kept alive during the extraction of lipids from it by using production [Source: FAO (1997)] mesoporous nanoparticles that extract oils from live cells of KNO3 con. the biomass to filtration and chemical extraction of oil from it can remaining pulp can be mixed with cyclo-hexane to extract the be avoided using the method which consists of treating algal remaining oil content.

wood and grass species.79 5 5 2. due to high content of carbohydrates/polysaccharides (Table 5) Gasification is a term that describes a chemical process by and thin cellulose walls. which are easier to be pyrolyzed Ethanol production: Algae are the optimal source for bioethanol to bio-oils and bio-gases.46 82 microalgae contain high content of cellular lipids. Saccharomycess cerevisiae is added when fuel and a chemical feedstock however under normal conditions the biomass starts degrading to initiate fermentation. Under dark anaerobic conditions. Other conversions are also The purification process of ethanol by distillation is an energy possible. types of energy fuels depending on temperature used for of conversion. This technology may be 28. Ethanol the technical potential of different biomass sources (marine production by dark fermentation in the marine green alga algae. condensable organic liquids.2 6. synthesis gas (syngas) by means of partial oxidation with air.09 2 2 2. often limited by the availability of nutrients. more suitable for microalgae because of the lower temperature ±2. effective. the cellular starch macroalgae exceed those of land plants. 1975). Compared to lignocellulose. bulky to handle. 4. 8. 4. resolvable Pyrolysis of biomass produces charcoal.56 30. Carbohydrate and Lipid content of some commonly candidates for ethanol production are Sargassum.e ethanol is drained from the tank and transportation fuel is limited. Ethanol. municipal solid waste) to be used Chlorococcum littorale was also investigated (Ueno et al.82 23 required for pyrolysis and the higher-quality of oils obtained 25. acetic acid. 2006).02 30. Generally two methods are employed which carbonaceous materials (hydrocarbon) are converted to a for the production of bioethanol from microalgal biomass.9 8. The starch of microalgae is higher alcohols. n. namely fermentation (biochemical process) and gasification oxygen and/or steam at high temperatures.76 34.. hydrogen and carbon dioxide were obtained as methane to methanol through photochemical conversion is fermentation products. typically in the range (thermo-chemical process) (Minowa and Sawayama. in energy farms.Mamta Awasthi and Rajiv Kumar Singh.03 1 8 ±2. The growth rates of marine starch was consumed within 24 h at 250C. 800-9000C.6 non-condensable gaseous products by a simple. A novel energy production system countries to produce ethanol from sugar crops and starch crops.9 Mean Ma Mi Mean M temperature. Pyrolysis is a phenomenon related to decomposition DW) of biomass under the condition of oxygen deficiency and high Min. 1999). Syngas can be burned directly or used as fuel for Fermentation is used commercially on a large scale in various diesel or gas turbine engines. Some prominent strains of algae that possible.12 9 wasteless and pollution free process. using microalgae with nitrogen cycling combined with low The biomass is grounded. 1995). The sugar is converted to ethanol by yeast. Gracilaria.6 11. acetone.. have a high carbohydrate content and hence. S. found algae 1984). Carbo Thermochemical conversion hydrat Thermal decomposition of algal biomass can yield different e (% Organism quadricauda Pleurotonium sp. and the starch is converted by temperature catalytic gasification of the microalgae has been enzymes to sugar. which have octane-enhancing properties of released from the cells with the aid of mechanical equipment or . ±SD ax. however. methanol and 27. The it is a gas and therefore. Chynoweth et al. ±5. growth is decomposition being accelerated at higher temperature. its use as a product of fermentation i. (1993) compared pumped to a holding tank to be fed to a distillation unit. polysaccharides and proteins. most notably a thermochernical conversion to produce intensive step (Minowa et al. and concluded that marine biomass offered the 1998).1 22. Methane is widely used as both a an enzyme and yeast. (Olson and Ingram. proposed (Sebnem Aslan. Anabaena sp.85± 9. ±3.29± 11. 27% of the cellular highest potential for biomethanation. 2011 can be converted to methane.33 14.2 25. promising Table 5. Euglena gracilis (Piorreck and Pohl. Lipid (% of DW) 20. Conversion of acetate. ±SD x. Prymnesium parvum.

most subjecting algae oil to hydroprocessing. 2010).Mamta Awasthi and Rajiv Kumar Singh. aviation fuel. profit and the design requires the combination and optimization Nevertheless.g. product separation. Integration approaches for pressure conditions in the presence of a catalyst and a solvent. the compounds. interesting. 2007). 1994). Bio-hydrogen production from micro algae has High content of water often exists in micro algae after been known for more than 65 years and was first observed in the harvesting which requires a great deal of energy to remove green alga Scenedesmus obliquus and in many other moisture from algal cells during pretreatment. Among the technologies used. Earlier.. of the total organic matters (Singh and Gu. Liquefaction has photosynthetic species (Boichenko and Hoffmann. its composition depends on the working of several factors such as biomass culture. transport of Hydrogenation saleable products and marketing. which could also be coupled to atmospheric CO2- Thermochemical liquefaction sequestration. The process consists of the addition of hydrogen conversion plants are located in or near the biomass growth atoms to the double bonds of a molecule through the use of a areas to minimize the cost of transporting biomass to the plants. owing to innovative technologies. Furthermore.. the thermochemical transport to conversion plants. must be established between the gaseous phase (hydrogen and Bio-refinery approach and other biofuels hydrocarbon phase). waste (liquid and solid) management. results addition of hydrogen (H2).. 1942). Integrated biodiesel production from microalgae (IBPCS) Thermochemical liquefaction requires temperature around 350 IBPCS enables the overall system to be operated at a and 395◦C in order to have the optimal amount of extracted oil. It was for photosynthesis research via an aerobic-anaerobic cycle reported that Dunaliella tortiolecta cells with 78. recycling. heat and mass. catalyst (Gaffron and Rubin. sustainable micro algal biodiesel production use hydrothermal Algal hydrogenation is a three-phase operation in which contact technology for direct liquefaction of algal biomass. Algal hydrogenation is of which all the non-fuel effluents are recycled to the growth performed by using an autoclave under high temperature and areas (Pushparaj et al. 2011 ethanol and methanol but pose fewer water solubility and phase promising and a potentially emission-free fuel stream for the separation problems. Liquefaction of petroleum distillation e. temperature. a model organism microalgae can provide hydrogen for hydrogenolysis. future. but at present. liquefaction seems to be more efficient than the extraction. growth management. while minimizing waste and pollution (Pushparaj et al. wet material can be used in the process. culturing of microalgae. algae biofuel content converts to oils directly. wet the green alga Chlamydomonas reinhardtii. .4% water developed by Melis and coworkers (2000). The yield of oils reached 37% projects were focused at obtaining biodiesel fuel. 1993). Moreover. it Algal biohydrogen production is recommended that a bio-refinery approach is the best solution Over the years microalgae for photo-biological to combine and integrate various processes to maximize hydrogen production from water are being developed into a economic and environmental benefits. 2010). In the idealized case. and solid particle phase (algal and catalyst) in order to improved by using the bio-refinery based production strategy achieve algal conversion and to promote the transfer of where all the components of the biomass raw material are used momentum. These factors can be simplified Hydrogenation is a reductive chemical reaction that and reduced to three main groups. which is obtained by algae uses moderate temperature and pressure and. 1993). Most been developed to produce bio-fuel directly without the need of studies on algal hydrogen production have been performed using drying micro algae (Singh and Gu. to produce useful products (Antoni et al. usually to saturate organic harvesting and processing of biomass. liquid phase (mixture of solvent and liquid The economics of biodiesel production can be significantly product). drying. braunii produced liquid oils at 57-64% interested in the possibility of obtaining other kinds of fuel from of dry weight under the conditions of a N2 pressure of 10 MPa at algae that are close in composition to fuel products obtained by 300°C in warm water and catalyzed by NaCO3. Dote et al. producers are becoming (1994) reported that B.

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