You are on page 1of 17


discussions, stats, and author profiles for this publication at:

The Techno-Economic Basis for Coproduct

Manufacturing To Enable Hydrocarbon Fuel
Production from Lignocellulosic Biomass

Article in ACS Sustainable Chemistry & Engineering April 2016

DOI: 10.1021/acssuschemeng.6b00243


2 98

11 authors, including:

Ling Tao Nancy Dowe

National Renewable Energy Laboratory National Renewable Energy Laboratory


Michael T Guarnieri Davinia Salvacha

National Renewable Energy Laboratory National Renewable Energy Laboratory


Some of the authors of this publication are also working on these related projects:

Engineering T. reesei for enhanced heterologous enzyme expression View project

Engineering T. reesei for enhanced heterologous enzyme expression View project

All in-text references underlined in blue are linked to publications on ResearchGate, Available from: Gregg T. Beckham
letting you access and read them immediately. Retrieved on: 28 October 2016
Research Article

The Techno-Economic Basis for Coproduct Manufacturing To Enable

Hydrocarbon Fuel Production from Lignocellulosic Biomass
Mary J. Biddy,* Ryan Davis, David Humbird, Ling Tao, Nancy Dowe, Michael T. Guarnieri,
Jerey G. Linger, Eric M. Karp, Davinia Salvachua, Derek R. Vardon, and Gregg T. Beckham*
National Bioenergy Center, National Renewable Energy Laboratory, 15013 Denver West Parkway, Golden, Colorado 80401, United
S Supporting Information

ABSTRACT: Biorenery process development relies on techno-economic

analysis (TEA) to identify primary cost drivers, prioritize research directions,
and mitigate technical risk for scale-up through development of detailed
process designs. Here, we conduct TEA of a model 2000 dry metric ton-per-
day lignocellulosic biorenery that employs a two-step pretreatment and
enzymatic hydrolysis to produce biomass-derived sugars, followed by
biological lipid production, lipid recovery, and catalytic hydrotreating to
produce renewable diesel blendstock (RDB). On the basis of projected near-
term technical feasibility of these steps, we predict that RDB could be
produced at a minimum fuel selling price (MFSP) of USD $9.55/gasoline-
gallon-equivalent (GGE), predicated on the need for improvements in the
lipid productivity and yield beyond current benchmark performance. This cost
is signicant given the limitations in scale and high costs for aerobic
cultivation of oleaginous microbes and subsequent lipid extraction/recovery.
In light of this predicted cost, we developed an alternative pathway which demonstrates that RDB costs could be substantially
reduced in the near term if upgradeable fractions of biomass, in this case hemicellulose-derived sugars, are diverted to coproducts
of sucient value and market size; here, we use succinic acid as an example coproduct. The coproduction model predicts an
MFSP of USD $5.28/GGE when leaving conversion and yield parameters unchanged for the fuel production pathway, leading to
a change in biorenery RDB capacity from 24 to 15 MM GGE/year and 0.13 MM tons of succinic acid per year. Additional
analysis demonstrates that beyond the near-term projections assumed in the models here, further reductions in the MFSP toward
$23/GGE (which would be competitive with fossil-based hydrocarbon fuels) are possible with additional transformational
improvements in the fuel and coproduct trains, especially in terms of carbon eciency to both fuels and coproducts, recovery and
purication of fuels and coproducts, and coproduct selection and price. Overall, this analysis documents potential economics for
both a hydrocarbon fuel and bioproduct process pathway and highlights prioritized research directions beyond the current
benchmark to enable hydrocarbon fuel production via an oleaginous microbial platform with simultaneous coproduct
manufacturing from lignocellulosic biomass.
KEYWORDS: Biochemicals, Lignocellulose, Integration, Biorenery, Biofuels

Lignocellulosic biomass oers signicant promise to oset a
worldwide fuel demand and has caused signicant concerns
regarding osetting food production for fuels.5
large portion of mankinds fossil fuel usage as part of a balanced As a means to ameliorate the concerns regarding the food
renewable energy portfolio for transportation fuels, renewable versus fuel debate and to further reduce greenhouse gas
chemicals, and robust materials, and as a partial source of emissions, support energy security, promote rural job develop-
sustainable power generation. Among all major renewable ment, and achieve fuel-scale production beyond starch- and
energy technologies examined to date, biomass is the primary sugar cane-based feedstocks, ethanol production from ligno-
source of renewable liquid fuels for vehicle, air, and maritime cellulosic biomass is under intense development at the
transportation.1,2 As highlighted in a number of studies, industrial scale. The current predominant route for lignocellu-
including the SCOPE Bioenergy and Sustainability study, well losic ethanol production proceeds via biochemical conversion
over 50 countries have biofuels mandates or blending targets.3 processes that conventionally includes a mild thermochemical
These mandates are dominated by ethanol, with typically pretreatment or fractionation step, enzymatic hydrolysis with
smaller biodiesel mandates, in the range between 1% and 20%
of the transportation pool.3,4 The vast majority of ethanol Received: February 2, 2016
production today comes from starch- or sugar cane-based Revised: April 27, 2016
industries, which exhibit a substantial scale limitation relative to

XXXX American Chemical Society A DOI: 10.1021/acssuschemeng.6b00243

ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

cellulolytic enzymes, and fermentation by an ethanologen such catalytically upgraded to a renewable diesel blendstock
as Saccharomyces cerevisiae or Zymomonas mobiliz.6,7 Certainly (RDB).41 The predicted RDB cost from this process is
many variations exist on that theme including, for example, $5.10/GGE in 2011 USD (hereafter the basis for cost gures
many dierent types of pretreatment8,9 and process cong- unless otherwise noted). The main cost drivers, and thus
urations with separate or combined hydrolysis and fermenta- identied research needs, associated with realizing a process
tion.10,11 Multiple techno-economic analysis (TEA) and life- based on this model include achieving FFA yield and
cycle analysis (LCA) studies have been conducted for various productivity of 0.28 g/g sugars and 1.3 g/L/hr, respectively,
congurations of industrial cellulosic ethanol plants, all of coupled to the ability to (1) secrete FFAs and (2) convert a
which point toward a path to produce ethanol that is at, or mixed sugar stream to FFAs. The productivity target was
close to, being competitive with petroleum-derived fuels with chosen at the time for consistency with what is achievable in
the potential ability to oset substantial greenhouse gas (GHG) ethanol fermentation,22 and the yield is within reach from
emissions,1225 and indeed, several cellulosic ethanol plants are literature values for FFA yields from glucose.41 However, we
currently being brought online worldwide. Going forward, note that both the previous productivity assumption and FFA
however, ethanol is a renewable additive primarily for gasoline- secretion have yet to be achieved to our knowledge in an
range fuels, but it is not directly applicable to a broader pool of oleaginous microbe. FFA secretion has been achieved in several
transportation fuels including diesel, jet, and maritime fuels nor nonoleaginous microbes, including the cyanobacteria Synecho-
does it typically suce as a complete substitute for gasoline in cystis sp. PCC6803,43 Escherichia coli,44 and Saccharomyces
most engines, a limit typically referred to as the blend wall. cerevisiae,4547 suggesting that fatty acid secretion is a distinct
Lastly, ethanol distribution does not follow the same channels possibility in oleaginous microbes. Moreover, the scale of
as petroleum-derived fuels. The issues of the limited fuel reactors considered previously did not account for the
market capacity, the need for another fuel distribution system, substantial dierences in aerobic and anaerobic bioreactor
and the ethanol blend wall have thus motivated research eorts cultivation of microbes. As demonstrated here, these
to move toward the production of drop-in infrastructure- assumptions have major eects on the estimated cost of an
compatible hydrocarbon fuels. oleaginous fuel platform.
Drop-in lignocellulosic biofuels beyond ethanol that are The cost projection of $5/GGE would represent a milestone
derived from biological26 or catalytic routes2734 have been toward the ultimate objective of producing hydrocarbon fuels at
examined primarily at the laboratory or pilot scale, with many prices concomitant with conventional petroleum fuels today
in the scientic literature rst focusing on the development of (e.g., $2$3/GGE).41 To achieve a lower price, signicant
processes starting from clean sugars. Many challenges remain process modications will likely need to be considered. One
in both biological and catalytic routes to hydrocarbon fuels, option to signicantly lower the cost of fuel production from
primarily related to realizing integrated processes that are able biomass, as is practiced throughout the petroleum rening
to completely convert lignocellulosic biomass into a nished industry, is to rely on the production of high-value coproducts
fuel. For example, inorganic components found in biomass, alongside fuels, as has been noted throughout the biomass
which are typically dicult to remove, oftentimes poison valorization literature.34,4859 Production of petrochemicals in
chemical catalysts.3537 Biological production of hydrocarbons many cases employs oxidation of hydrocarbon platform
from biomass-derived sugars typically suers from the molecules, and relative to petroleum, biomass-derived platform
challenges of insucient titers, productivities, and yields molecules from polysaccharides and lignin already exhibit
when a pathway is rst introduced as well as toxicity eects signicant oxygen content, making them, from a stoichiometric
stemming from inhibitors produced during biomass decon- perspective, more attractive building blocks for oxygen-
struction.3840 These challenges, among many others in process containing chemicals if appropriate molecular transformations
integration and yield improvements, must be overcome to cost can be realized.53,59 The production of biobased chemicals,
eectively produce hydrocarbon biofuels from lignocellulosic however, also presents a substantial set of technical challenges.
biomass. For example, signicant emphasis has been placed in pioneering
To bring hydrocarbon fuel production to a level of reports on the production of single-platform molecules, often
technological viability similar to that of ethanol production, from sugars for uses such as chemical and polymer
TEA and LCA studies will be key to consistently evaluate and precursors.48,50,53 Especially for polymer and specialty chemical
compare the myriad process options that exist and are being applications, precursor purity is often of utmost importance,
constantly proposed and developed. To that end, Davis et al. which in an integrated process from lignocellulosic biomass will
recently published a peer-reviewed publically available report require scalable cost-eective separations and cleanup strat-
describing a projected TEA and LCA model that examines the egies. As a result, it has also been suggested that biobased
cost of producing hydrocarbon biofuels via a biological- chemicals should rely on the intrinsic heterogeneity and
upgrading step from lignocellulosic sugars and sustainability intrinsic polymeric structure already present in biomass to
considerations within the bioenery conversion facility.41 The make functional replacement materials rather than depolyme-
primary purpose of this model was to identify research gaps and rization to monomers that can then go on to be converted to
cost drivers to achieve a near-term 2017 technical target to building blocks.56 Regardless, whether biomass-derived sub-
produce hydrocarbon fuels at a cost of USD $5 per gasoline strates are converted to new platform molecules or used
gallon equivalent ($5/GGE). The modeled biochemical directly as polymers, process integration issues, including
conversion process described in that work employs a two- separations, cleanup, and quality of the nal products, will be
step pretreatment process consisting of a deacetylation step of paramount importance to replace products used by
followed by dilute-acid pretreatment,42 enzymatic hydrolysis consumers currently derived from petroleum. Moreover, the
with cellulase enzymes, and an aerobic cultivation step to market size disparity between fuels and chemicals will be a key
employ an oleaginous microbe to produce free fatty acids driver in the attainable scale for a single process, further
(FFAs), which are secreted into the broth, recovered, and warranting the need to develop modular processes that can be
B DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

Figure 1. Overview of design block ow diagrams from (A) baseline fuels only design and (B) coproduction of fuels and chemicals design.

adjusted to produce several dierent chemicals, either spread after dilute-acid pretreatment, and the cellulose-derived sugars
across multiple bioreneries, or within the same biorenery, are used to produce lipids for RDB. Succinic acid is chosen as a
depending on the economies of scale for a given process representative molecule to demonstrate the potential for
conguration or coproduct. Overall, for the production of coproduct pathways within this framework for an integrated
hydrocarbon biofuels from lignocellulose, the basis for biorenery, with preliminary research demonstrating biological
manufacturing coproducts alongside fuels must be quantied production of succinic acid at encouraging performance levels.
with TEA to ascertain the relative contributions that coproducts When viewed as an intermediate molecule, there is a strong
can make to the overall economic viability. potential for a greater demand of succinic acid given that there
To that end, here we present TEA models for several process are numerous known technologies for the production of a wide
scenarios centering on the core concept of a biorenery slate of high-value large market commodity chemicals from
wherein hydrocarbon fuels and chemicals are produced in succinic acid.48,53,60
parallel from lignocellulosic biomass similar to the simultaneous The model predicts that the production price of hydrocarbon
production of fuels and chemicals practiced in petroleum
fuels can be signicantly improved by the simultaneous
rening. The baseline process considers economics of fuel
production of chemicals from other biomass fractions. Key
production alone similar to that noted above as presented in
Davis et al. and is a two-step pretreatment approach of ndings from this work regarding process selection are as
enzymatic hydrolysis and biological production of lipids and a follows: (1) Maximizing carbon eciency, especially to higher
catalytic step to produce RDB from both hemicellulose and value coproducts, is paramount. (2) Recovery and purication
cellulose-derived sugars.41 However, for this case, a number of of both fuels and coproducts are major cost drivers that will
updates are made to reect more realistic experimental benet signicantly from research and development activities to
benchmarks (such as intracellular lipid productivity metrics improve applicability at the biorenery scale. (3) Product
via oleaginous yeasts rather than more idealized high- selection and price will greatly inuence the process exibility
productivity FFA secretion) and to incorporate improved and the ability to balance process severity. Overall, this study
design details (such as the use of revised design/cost estimates quanties the economic baseline for the manufacturing of next-
for bubble column bioreactors). We then examine a coproduct generation hydrocarbon fuel bioreneries, demonstrates that
manufacturing scenario wherein succinic acid is produced coproducts can enable the economics of hydrocarbon fuel
predominantly from hemicellulose-derived sugars fractionated production, and provides a framework for further research and
C DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

development in a next-generation integrated lignocellulosic feedstock cost. The biomass is conveyed to the pretreatment
biorenery. area where it is rst processed in a deacetylation step that uses a

sodium hydroxide soaking process (loaded at 17 mg of NaOH/

SUMMARY OF MODEL g of dry biomass) at 80 C for 1 h in a set of parallel batch
reactors. This process step has been shown to remove various
As illustrated in Figure 1A, the base case design produces components of the biomass that are either potentially toxic in
hydrolysate via deacetylation and dilute-acid pretreatment of downstream biological upgrading or inert biomass components,
biomass followed by enzymatic hydrolysis. Unconverted including acetate, lignin, extractives and ash, and improves
biomass solids (mostly lignin) are removed from the hydro- overall yields and reduces capital cost due to lower process
lysate through a occulant-aided solid/liquid separation, and throughputs.41,42,61 The solubilized fraction of the biomass is
the remaining liquor fraction is concentrated prior to biological drained from the batch reactors and sent to wastewater
upgrading. The hydrolysate, which is rich in both C5 and C6 treatment. Overall, this liquor fraction consists of a range of
sugars, is biologically converted in an aerobic process to lipids biomass components with solubilization and removal of 100 wt
that are stored intracellularly by an oleaginous microbe. The % of the extractives, 75 wt % of the ash, 20 wt % of the lignin, 2
lipid fraction is extracted from the cells in a wet extraction wt % of the xylan, 50 wt % of the sucrose, and 88 wt % of the
process and subsequently upgraded in a hydrotreating unit to acetate. The conversion in the deacetylation step is consistent
produce RDB. Supporting processes, including wastewater for all of the cases studied.
treatment, utilities (including cooling water, chilled water plant, The remaining solids are conveyed to the dilute-acid
instrument air, and process water), and the combined heat and pretreatment reactor at a total solids loading of 30 wt %,
power system (consisting combustor, boiler, and turbogener- where a large fraction of the hemicellulose carbohydrates are
ator) are also incorporated in the design and economics. converted to soluble sugars including xylose, arabinose,
A variation of the process design, as outlined in Figure 1B, galactose, mannose, and some glucose. The basis for the
considers the conversion of a separate C5-rich hydrolysate design of the reactor system is described in a previous study.41
stream to a chemical coproduct. In this case, an additional The biomass is treated with 9 mg of H2SO4/g dry biomass
solid/liquid separation step is introduced after the pretreatment (based on acid loading present in the pretreatment reactor),
process, and the C5-rich liquor is sent to a biological upgrading with a total residence time in the reactor of 5 min at a
process for continuous conversion to succinic acid, followed by temperature of 165 C. The overall conversion reactions are
the required separation and purication steps. The solids (C6- provided in Table 2 and summarize both the current
rich hydrolysate stream) continue to enzymatic hydrolysis and benchmark case as demonstrated in National Renewable
follow the same process steps to RDB as outlined in the base Energy Laboratory pilot plant runs and a projected improved
design. case.61 Following the acid pretreatment, the hydrolysate whole
Additional details of the design and the basis for both the slurry containing 30 wt % total solids and 16 wt % insoluble
current experimental benchmark and an improved case are solids is neutralized with ammonia in stoichiometric quantities
summarized in the following subsections. The base case process to raise the hydrolysate pH to 5, and the stream is diluted with
(whole hydrolysate to fuels alone) is discussed rst, followed by water to a total solids concentration of 20 wt % prior to
the modications made for the alternative coproduction of enzymatic hydrolysis.
chemicals and fuels process (C6-to-fuels and C5-to-copro- Enzymatic Hydrolysis. In the base case, the whole slurry
ducts). hydrolysate stream is fed at a 20 wt % total solids to a high-
Pretreatment. Milled biomass, with a composition solids continuous reactor for enzymatic hydrolysis by a cellulase
described in Table 1, enters the process facility and is assumed enzyme prepared on-site. The partially hydrolyzed slurry is next
to be throat-of-reactor ready, i.e., any preprocessing required batched to one of several parallel bioreactors. Hydrolysis is
prior to the rst unit operation is included in the delivered completed in the batch reactor with an overall hydrolysis time
of 3.5 days in the continuous and batch steps. This process
Table 1. Delivered Feedstock Composition Assumed in converts both glucan and residual xylan in the solids to
Process Design monomeric glucose and xylose, respectively. Table 2 summa-
component composition (dry wt %) rizes the key process conditions and yields for both the
glucan 35.05
benchmark and improved cases for the enzymatic hydrolysis
xylan 19.53 process.
lignin 15.76 Hydrolysate Conditioning: S/L Separations and
ash 4.93 Concentration. The slurry is further conditioned for down-
acetatea 1.81 stream biological upgrading by removal of insoluble solids
protein 3.10 required for eective aerobic biological conversion and
extractives 14.65 subsequent product purication. A vacuum lter press is used
arabinan 2.38 for this solid/liquid separation step. To facilitate and improve
galactan 1.43 ltration, polymer occulants at a loading of 10 g/kg insoluble
mannan 0.60 solids are added to reduce the overall required lter area. The
sucrose 0.77 process conditions for both the benchmark and improved cases
total structural carbohydrate 58.99 are outlined in Table 2 for this step, with the benchmark values
total structural carbohydrate + sucrose 59.76 based on data demonstrated from bench scale studies reviewed
moisture (bulk wt %) 20.0 in Sievers et al.62 The solids removed in the lter press are sent
to the combustor/steam turbogenerator, which provides all
a process heat and a portion of the power requirements for the
Represents acetate groups present in the hemicellulose polymer;
converted to acetic acid in pretreatment. integrated process design.
D DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

Table 2. Overall Yields for Benchmark and Improved Case (Biomass Deconstruction and Fuel Production Operations)
experimental benchmark case improved case
solids loading (%) 30% 30%
acid loading (mg H2SO4/g dry biomass present in reactor) 9 9
% Conversion: (Glucan)n + nH2O nGlucose 10% 10%
% Conversion: (Xylan)n + nH2O nXylose 76% 78%
% Conversion: (Galactan)n + nH2O nGalactose 90% 90%
% Conversion: (Arabinan)n + nH2O nArabinose 90% 90%
% Conversion: Acetate Acetic Acid 100% 100%
% Conversion: Lignin soluble lignin 5% 5%
S/L separation:soluble sugar loss to solids 4% 1%
enzymatic hydrolysis + hydrolysate conditioning
enzyme loading (mg/g cellulose) 12 10
enzymatic hydrolysis time (days) 5.0 3.5
% Conversion: (Glucan)n + nH2O nGlucose 86% 90%
% Conversion: (Xylan)n + nH2O nXylose 93% 93%
S/L separation: soluble sugar loss to solids 5% 5%
bioconversion (lipid production)
lipid productivity (g/L/hr) 0.34 0.4
lipid content (wt %) 60% 70%
% Conversion: Glucose Lipid [total glucose utilization]a 75% [100%] 82% [100%]
% Conversion: Xylose Lipid [total xylose utilization]a 44% [59%] 80% [98%]
bioconversion metabolic yield [process yield] (g/g)b 0.25 [0.24] 0.27 [0.27]
lipid recovery + upgrading
extraction pretreatment acid loading (wt % of feed liquor) 1% 1%
extraction solvent loading (g hexane/g cell mass) 5 5
extraction net lipid recovery 90% 90%
hydrotreating fuel yield (g RDB + naphtha/g lipid) 0.81 0.81
hydrotreating net H2 makeup demand (wt % lipid feed) 1.8% 1.8%
First number represents sugar conversion to desired product (lipids); values in brackets indicate total sugar utilization (including diversions to cell
mass). bMetabolic yield = g product per g sugars converted; process yield = g product per g total sugars available for bioconversion.

The claried soluble sugar stream is split with roughly 15% the target lipid productivity rate (g/L/hr) and the associated
routed directly to bioconversion to initiate fed-batch oxygen uptake rate (OUR), which is set in the model to be
fermentation, and the majority (85%) concentrated from 20 equal to the oxygen transfer rate (OTR). The required OTR is
to 50 wt % total sugars in a mechanical vapor recompression achieved in the model by varying the aeration rate to the bubble
(MVR) evaporation system. The concentrated sugar stream columns, given a xed vessel geometry and air compressor
exiting the evaporators is cooled and fed to the bioreactors. pressure (liquid height plus hydraulic losses). Further details on
Biological Conversion. The biological conversion step vessel design and operation, as well as underlying drivers
consists of a seed train of increasingly larger stirred tank behind the OTR/OUR in the modeled system, are provided in
reactors in series that are dedicated to biomass seed the Supporting Information. Table 2 provides a summary of key
propagation (grown with a 10% split of the starting available conversion and yield parameters for the bioconversion step
hydrolysate sugars) and a system of aerobic bubble columns for attributed to the experimental benchmark and projected
the production of lipids. Many experimental eorts in the improved cases.
literature have focused on the development and evaluation of Lipid Separations. The hydrocarbon intermediate product
intracellular lipid accumulation in model organisms such as
is composed of intracellular lipids (primarily triglycerides). The
Saccharomyces cerevisiae63 or Escherichia coli6466 or oleaginous
lipid fraction is extracted from the cells in a wet extraction
microbes such as Yarrowia lipolytica,6769 Rhodosporidium
process. The extraction step follows a similar schematic as
toruloides,7072 and Lipomyces starkeyi,7274 among many
others. The biomass composition in the current model is published for extraction of lipids from algal biomass with
based on L. starkeyi.74 The production bioreactors are modeled demonstrated yields up to 92.5%,75,76 consisting of a dilute-acid
as bubble columns each with 500 m3 (500,000 L) of total treatment operation targeting 1% acid loading at 150 C,
internal volume operated in a fed-batch mode of operation, followed by cooling, ashing, and neutralization (all steps being
with an initial 50% ll level up to a nal 80% ll level (400 m3 similar to the initial biomass pretreatment step discussed
maximum ungassed working volume) over the course of the previously). The material is then routed to a countercurrent
fed-batch cycle. The reactors are cooled through an external extraction column with hexane solvent at a total 5:1 solvent
pump-around loop circulated through a chilled water heat loading (solvent versus yeast dry cell weight). The extract
exchanger. consisting of primarily solvent and lipids is sent to a stripping
Key cost and design considerations for this system are column to recover the hexane, while the aqueous ranate
attributed to the aerobic nature of the bioconversion step, with product is sent to wastewater treatment. The key process
submerged aerobic production of the yeast cells dependent on conditions and yields for this step are summarized in Table 2.
E DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

Upgrading and Recovery. Finally, the recovered lipid therefore reduced to 17.5 wt % for this modied design.
product exiting the extraction/separation step is upgraded via However, the downstream yields and all process design
hydroprocessing to a hydrocarbon product, primarily RDB with assumptions for the enzymatic hydrolysis and conversion/
a small naphtha coproduct. This operation is designed for upgrading steps are consistent with the fuel-only base case.
deoxygenation and saturation of the lipid material, yielding a The C5-rich hydrolysate liquor continues to a parallel
paranic end product suitable for blending. The process biological upgrading train for chemical coproduction. For this
conditions and yields for this step are summarized in Table 2 design, succinic acid has been identied as an example target
and are based on detailed experimental data documented in chemical product, utilizing a process modeled with Actino-
Marker et al.77 for the upgrading of vegetable oils. The lipid bacillus succinogenes as the production organism. This organism
product is modeled as a representative triglyceride component can utilize pentose and hexose sugars, as well as hydrolysates,
consisting of oleic (18:1), stearic (18:0), and palmitic (16:0) with the production of succinic acid being greatly enhanced by
acid chains, which constitute the majority of the fatty acid the presence of CO2.80,81 Succinic acid is meant to serve as a
prole characterized in many oleaginous yeast.74 On the basis representative molecule to demonstrate the potential enabled
of this representative component, plus the targeted 81 wt % fuel by introducing coproduct pathways within this integrated
yield (79% RDB plus 2% naphtha versus lipid feed), the biorenery framework. This integrated biorenery concept
modeled hydrogen consumption is roughly 1.8 wt % of the lipid should thus be applicable to the coproduction of fuels and a
feed to the upgrading step. Final fuel blendstock yields wide of array of other chemical intermediates.
following the upgrading section are calculated based on the In this design, the C5 liquor is initially split, with 10% of the
total energy yield and are converted to a GGE basis according liquor being sent to the seed fermentor used to grow A.
to lower heating values (LHV), assuming standard published succinogenes. The other 90% of the liquor is sent to the
LHV values for gasoline.78 O-gases from the upgrading section continuous biological reactor system. CO2 is also fed to the
are combined and routed to the boiler, while produced water is bioreactor at a rate of 0.1 VVM, which is adopted from
sent to wastewater treatment. literature.82 Sodium hydroxide is added to the biological
Modications to the Base Design for Coproduction of conversion process to maintain the pH of the system near 7,
Fuels and Chemicals. While the base case design is focused with stoichiometric amounts of NaOH added to the
solely on the production of fuels in the biorenery, an fermentation broth. The near-term conversion for the
alternative scenario is investigated that considers the economic bioreactor for the improved case are set at 0.73 g succinic
impact of the coproduction of chemicals and fuels. For this new acid/g pentose sugars (xylose and arabinose) and 0.73 g
basis with coproduction, the benchmark design is modied to succinic acid/g hexose sugars (glucose and galactose), with
separate and upgrade the C5-rich hydrolysate stream to pentose sugar utilization targeted to improve to similar levels as
chemicals via biological conversion, while the residual C6-rich those reported publicly for hexose sugars.83 Selectivity to acid
slurry is upgraded to fuels. byproducts is assumed to be minimized to 0.078 g/g C5 sugars
More specically, the hydrolysate slurry after dilute-acid and 0.081 g/g C6 sugars for acetic acid coproduction and 0.02
pretreatment is immediately processed in a vacuum lter to g/g C5 sugars and 0.03 g/g C6 sugars for formic acid
separate solubilized hemicellulose sugars from cellulose and coproduction.83 The succinic acid productivity rate is targeted
residual unconverted solids. The current benchmark case for to be 2.0 g/L/hr, which is realistic for the continuous
this solid/liquid separation step is based on bench-scale fermentation pursued for this design. Recent results have
demonstrations on corn stover-derived pretreated hydrolysate. demonstrated that succinic acid can be produced from corn
It was demonstrated that recoveries of 96 wt % of the sugars stover hydrolysates at productivities of 1.45 g/L/hr utilizing a
using a wash ratio of 12.4 L/kg (wash water versus liquor in the continuous fermentation process and overall metabolic yields at
lter cake) and a lter capacity of 15.8 kg insoluble solids (IS)/ 0.78 g succinate/g of sugar consumed.84 These metrics are the
m2 h could be achieved. The target case utilizes process values incorporated for the experimental benchmark case.
assumptions based on previously reported experimental results Since A. succinogenes produces not only succinic acid but also
outlined in Sievers et al.79 In that study, a hydrolysate produced acetic and formic acids as byproducts, additional processing is
by dilute-acid pretreatment, but without deacetylation, was required to purify the succinic acid from the other organic
separated with a sugar recovery eciency of 99 wt % utilizing a acids, as well as from salts and other contaminant components
wash ratio of 4.3 L/kg and lter capacity of 20 kg IS/m2 h. present in the starting hydrolysate. The design basis for the
The recovered solids that are rich in unconverted cellulose succinic acid purication operation is based on recent patents
are sent to the enzymatic hydrolysis step and further upgraded by Myriant and Novasep for the recovery of succinic acid.85
to fuels, whereas the liquid, which will be referred to as the C5- The whole broth is rst ltered through an ultraltration step
rich liquor, is sent to the C5 conversion train for the production to remove cells and large insoluble particles from the broth.
of chemicals. The solids fraction is routed to enzymatic These recovered solids are sent to the boiler. The cleaned broth
hydrolysis at a 17.5 wt % total solids loading, which diers from is then introduced to a simulated moving bed unit (SMB-ICX),
the concentration commonly employed in whole slurry which converts the sodium succinate to succinic acid via ion
processing and that was discussed earlier for the base case. exchange. The succinic acid-rich stream is then further puried
Since the coproduction of chemicals and fuels pathway employs through nanoltration to help remove any residual solids and
a washed solids hydrolysis whereby a large fraction of soluble impurities that might color the succinic acid crystals. The dilute
solids (namely, hemicellulose sugars) are removed, the overall succinic acid stream is then further concentrated in an
viscosity of this stream is increased due to the higher fraction of evaporation process and then crystallized. The succinic acid
insoluble solids relative to total solids. As the material crystals are recovered via a centrifugation unit, dried via a belt
approaches 20% IS, it becomes considerably more dicult to conveyor, and sent to storage. Approximately 90% of the
pump and agitate in the early stages of hydrolysis, often mother liquor from the crystallizer, which contains nearly all of
resulting in reduced glucose yields. The total solids loading is the unconverted sugar, is recycled back to the fermentation
F DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

reactor for further conversion. The overall succinic acid improved cases was determined for both process pathways
recovery eciency is set to 82% with the bulk of the succinic and is summarized in Table 4. The overall MFSP for the fuel-
acid losses occurring in the crystallization process.
Economic Analysis Details. Utilizing the conceptual Table 4. Comparison of Economics for Two Scenarios
process design described above, Aspen Plus process models Considered (Improved Projection Cases)
were developed to estimate key material and energy ows in
the process. The overall variable and xed operating costs, as base case
coproduction of
fuels and
well as the capital costs, were then estimated for the integrated (fuels only) chemicals
design and the calculated ow rates. The capital costs, which MFSP ($/GGE) $9.55 $5.28
are outlined in Tables S3 and S4, are based on vendor designs annual fuel yield (MM GGE/yr) 24.1 15.0
and cost estimates for the major processing equipment. The chemical coproduct yield (MM tons/yr) 0 0.127
details of these equipment designs have been published in total capital investment (MM$) 921 1216
Humbird et al. and Davis et al. design reports.22,41 The cost total variable operating cost (MM$/yr) 89 135
associated with the recovery and purication stages in the coproduct credit (MM$/yr)a 6 247
design for chemicals production was taken from recent design total xed operating cost (MM$/yr) 21 27
estimates from external vendors. Raw material prices were a
The coproduct in the fuels-only model is electricity, while for the
drawn from a broad range of sources as reviewed in Table S5. fuels/chemicals case it is succinic acid.
A discounted cash ow analysis was used to determine the
overall minimum fuel selling price (MFSP) of the process
under a specic set of nancial assumptions. The nancial only base case is $9.55/GGE with a total capital investment of
assumptions used to calculate the MFSP are reviewed in Table $920 MM. The cost breakdown for the major process areas is
3 based on a mature nth plant and consistent with prior provided in Figure 2A. Roughly 39% of the capital cost is
attributed to the biological conversion section, which requires
Table 3. Financial Assumptions and Design Basis 97 bubble column reactors to process the full hydrolysate
plant life 30 years stream based on the projected lipid productivity of 0.4 g/L/hr.
plant throughput 2000 dry metric tons/day biomass This translates to intensive capital expenditures for this section
cost year dollar 2011 dollars of the process and leads to the greatest contribution to the fuel
capacity factor 90% price, accounting for 31% of the MFSP. The annual total
discount rate 10% variable cost of the process is $84 MM/year. Biomass feedstock
general plant depreciation 200% declining balance (DB) contributes the most to this value at $57.5 MM/year,
general plant recovery period 7 years accounting for roughly 25% of the MFSP. The second largest
steam plant depreciation 150% DB raw material cost occurs in the lipid extraction section, in which
steam plant recovery period 20 years the demand for sulfuric acid, hexane, and ammonia translates to
federal tax rate 35% an annual cost of $8.2 MM/year or 3.5% of the MFSP, while
nancing 40% equity the hydrogen required for fuel nishing accounts for 1% of the
loan terms 10 year loan at 8% APR MFSP. Enzymes represent a signicant driver accounting for
construction period 3 years 5% of the MFSP, even at the low loadings of 10 mg/g of
rst 12 months expenditures 8% cellulose projected in the improved design. The total variable
next 12 months expenditures 60% cost also includes a credit of $6 MM/yr associated with the sale
last 12 months expenditures 32% of excess electricity to the grid. The primary power consumer in
working capital 5% of xed capital investment the process is the bioconversion section, which requires 36% of
start-up time 6 months total biorenery electricity demand, attributed to aeration
revenues during start-up 50% requirements. Overall xed costs, which are partially a function
variable costs during start-up 75% of capital costs, are $21.2 MM/yr.
xed costs during start-up 100% Economics of Coproduction of Fuels and Chemicals.
When the base case is modied and the C5-rich stream is
separated from the unconverted biomass prior to enzymatic
published work.41,8688 All coproducts were treated using a hydrolysis and used to produce chemicals rather than fuel, the
market value allocation approach in a consistent manner with overall MFSP of the process is decreased by 45% to $5.28/
previously published studies, in which a credit is accounted for GGE in the projected improved case. Since this modied
in the operating costs and is allocated based on the coproduct design diverts some of the sugar stream originally available for
market value.41,8688 On a relative mass yield basis, a higher fuel production, the overall fuel production rates are reduced
yield of the chemical coproduct is observed relative to the fuel from 24 MM GGE/yr in the baseline case to 15 MM GGE/yr
yield for a number of the cases considered in this study. in the modied design. The addition of the C5 upgrading
Alternative strategies could be utilized to estimate the cost of strategy also results in an increase in the overall capital cost of
the chemical coproduct rather than the fuel if the fuel is the process, with the installed cost of this added process train
assigned a specic price. However, the primary focus of this being just below $225 MM or 32% of the overall capital cost of
study is to understand the cost of fuel production, and this the facility. There are two major areas that dominate this
evaluation is outside of the scope of this study.

additional capital cost. Recovery and purication of the succinic

acid product accounts for $134 MM, while the additional
RESULTS solidliquid separation processes add $45 MM and account for
Fuel-Only Base Case Economics. Utilizing the design and 21% of the capital costs of the C5 upgrading section. The
economic basis described above, an MFSP for the projected remaining 15% of the cost is the result of the continuous
G DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

Figure 2. Cost breakout by process area for two scenarios considered (improved projection cases): (A) base case fuel-only design and (B)
coproduction of fuels and chemicals design.

Figure 3. Sensitivity analysis results for the coproduction of fuels and chemicals process design (improved case, $5.28/GGE MFSP basis).

biological reactor design and the auxiliary equipment required total, however, there remains a 30% increase in the total facility
for succinic acid fermentation. These additional costs of the C5 capital investment to $1,216 MM for this process scenario.
The addition of the C5 upgrading train also increases the
upgrading section are partially oset by reduced overall
variable operating cost of the process by 50% when excluding
throughput in the C6 fuel train, which decreases the number any coproduct credits. The process requires $42 MM/yr for
of bubble column reactors to 60 and reduces the installed raw materials required for product separations and purication
capital cost of the biological upgrading section by $30 MM. In with nearly 90% of this total cost being attributed to
H DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

regeneration of the resin (sulfuric acid and caustic) utilized in that have been invested throughout the process up to this nal
the recovery design and the caustic required in biological step, where it is critical to recover as much product as possible.
conversion. Beyond this, the process now requires an electricity In the case of recovery and purication of the chemical
import of 11 MW at a cost of $6.1 MM/yr due to the coproduct, the cost of the recovery section accounts for over
introduction of a recovery and purication process that is 65% of the capital cost of the chemical production train and has
power intensive demanding 990 kWh/ton of succinic acid a signicant impact on the overall MFSP of the process. Indeed,
produced. This import is on the same order of magnitude as the when the capital cost of the coproduct recovery and purication
export estimated in the baseline design. Due to the higher section is modied by 25%, the MFSP changes by $0.68/GGE.
capital cost and increased complexity of the modied design, These results highlight that both fuel and chemical coproduct
the overall xed operating cost of this scenario was $27 MM/yr. recovery processes are paramount to controlling costs for this
Sensitivity Analysis for Coproduction of Fuels and biorenery scenario.
Chemicals. A number of the underlying process design For recovery of sugar intermediates, the solidliquid
parameters projected in the improved case have a signicant separation following pretreatment is also a key cost driver as
impact on the economics of the overall integrated design. As it impacts succinic acid yields, while separations after enzymatic
highlighted in the single-point sensitivity analyses summarized hydrolysis is a driver for fuels production. For the separation
in Figure 3 for the coproduction of fuels and chemicals, cost performed after pretreatment, reducing the eciency from the
drivers exist throughout the process design. Therefore, it is improved target value of 99% to the experimental baseline of
critical to understand the impact that each of these drivers has 96% increases the MFSP by $0.36/GGE, whereas 100%
on improving the MFSP and to prioritize further research. In separation eciency reduces the MFSP by $0.11/GGE. As
particular, sugar production and conversion are major drivers noted previously, this solid/liquid separation step represents
throughout the process. In the biomass pretreatment section, if 21% of the C5 upgrading capital cost, and if the cost for this
xylose yields could be increased from the target value of operation increased or decreased by 50% (also a measure of
78.5%80%, the MFSP would be reduced by $0.15/GGE due ltration capacity), then the MFSP would change by $0.45/
to the higher amount of xylose available for the production of GGE. The biggest driver in this section is the wash ratio
succinic acid. Since the succinic acid biological conversion can utilized. If the wash ratio were increased by 50%, then the
utilize minor sugars, an increase in arabinose production by 5% MFSP would be increased by $0.52/GGE due to the higher
would further decrease the MFSP by $0.06. In the enzymatic throughput of water sent to the C5 train and the additional
hydrolysis step, conversion to glucose also has a major impact; capital required to process and purify the diluted stream.
an increase in glucose yield from 90% to 95% reduces the Similar to the solidliquid separation immediately following
MFSP by $0.23/GGE, while a decrease in glucose yield to 86% pretreatment, for the solidliquid separation process after
results in an increase in the MFSP by $0.11/GGE. enzymatic hydrolysis, the lter costs, wash ratio, and recovery
When considering conversion eciency of the sugars to the eciencies are major drivers; however, their impact on costs is
desired product, glucose conversion is a major driver in the varied. The lter cost has the largest impact, as modifying this
lipid production train as lowering the yield by 20% would parameter by 50% will result in a $0.19/GGE impact on the
increase the MFSP by $0.49/GGE. In comparison, a similar MFSP. The impact on the MFSP is diminished in this section
reduction in xylose conversion in the lipid train only increases relative to this parameter for the unit following pretreatment, as
the MFSP by $0.05/GGE due to signicantly lower levels of the absolute lter capital costs are roughly 60% lower due to
xylose available for conversion in this train. In the succinic acid the reduced throughput in this portion of the process.
train, decreasing the conversion of xylose by 20% would Variations in the wash ratio also have less of an impact on
increase the MFSP by $0.75/GGE. This change is much higher costs (variations of 50% result in $0.16/GGE change to the
than the impact from glucose on lipid production since succinic MFSP), owing to the use of a concentration unit immediately
acid is substantially more valuable than the fuel on a mass basis. downstream of this lter step, thus a more dilute hydrolysate
Glucose conversion in the succinic acid train also impacts the stream in this case merely impacts the cost of concentration
MFSP, despite the lower level of glucose in this train, with a rather than the entire processing train. Yield impacts are also
20% reduction increasing the MFSP by $0.23/GGE. Although smaller, such that a 5% reduction in sugar yield increases the
the minor sugar concentration is low, a reduction in conversion MFSP by $0.08/GGE. Finally, varying the occulant loading
still has a signicant economic impact. For example, a reduction requirements by 50% aects the MFSP by $0.09/GGE.
in arabinose conversion by 20% increases the MFSP by $0.11/ Raw material prices are also key drivers throughout the
GGE. process. Changes of 50% in the amount of acid required in
Ecient recovery of the nal products, as well as the sugar pretreatment impacts the MFSP by $0.08/GGE due to both the
intermediates, at the lowest cost possible is also a key driver in change in acid required as well as the change in ammonia
the overall cost of the process. For the case of lipid recovery, required for neutralization. In the enzymatic hydrolysis section,
the cost of extraction and recovery accounts for roughly 20% of enzyme loading is a more important driver. Increasing the
the overall MFSP. If the lipids could hypothetically be required enzyme loading from 10 mg/g of cellulose to the
recovered from cells triggered to autolyse, the extraction and current benchmark value of 12 or even higher to 20 mg/g
recovery process could be eliminated, and assuming consistent loading increases the MFSP by $0.12/GGE and $0.78/GGE,
recovery eciency with the standard extraction basis, the MFSP respectively.
would be lowered signicantly by $1.10/GGE. Alternatively, The primary biological metrics dictating economics in the C6
making use of the extraction operation for lipid recovery, even fuel train are achievable lipid productivity and lipid content of
small deviations from the target lipid extraction eciency the cell. As shown in the tornado plot in Figure 3, productivity
translate to signicant MFSP impacts, e.g., $0.43/GGE has one of the biggest impacts on the MFSP of the entire
attributed to a 7% deviation from the targeted 90% recovery; process, particularly at lower values. Given the large capital
this is due to the signicant capital and operating expenditures costs of the aerobic bioreactors in the C6 train, increasing the
I DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

productivity from 0.4 to 0.6 g/L/hr would decrease the MFSP It is also important to highlight that the assumed price of the
by $0.53/GGE, while decreasing the productivity to 0.2 g/L/hr coproduct carries a dramatic eect on the overall MFSP of the
would increase the MFSP dramatically by $1.54/GGE. In light process. If the coproduct value were decreased from $0.97/lb to
of this strong nonlinear sensitivity, the impact of productivity is $0.75/lb, then the 0.4 g/L/hr productivity basis would increase
explored further in Figure 4A, which expands to consider a the MFSP by more than 70% to $9.04/GGE. In contrast, if the
coproduct was valued at a higher price of $1.20/lb, then the
MFSP for the 0.4 g/L/hr basis would decrease to $1.42/GGE.
Similar trends are seen in the lipid content curve as well (Figure
4B), highlighting the dramatic eect of coproduct price on the
One additional scenario was considered for the coproduction
process. Since both the C5 and C6 trains can convert C5 and
C6 sugars to the desired products, the design was modied to
consider splitting the sugar stream after enzymatic hydrolysis.
This approach not only allows for the expensive solidliquid
separation process after pretreatment to be eliminated but also
allows for exibility in the amount of sugar that is converted to
fuels versus chemicals, as might be dictated by the relative value
of each fuel or product or other dynamic constraints imposed
on the biorenery. Further, we estimate that the removal of the
solidliquid separations process reduces the cost of the
concentrated sugar intermediate price from $0.20/lb of sugar
for the split case to $0.18/lb of sugar.
The impact of this design modication on the process
economics is illustrated in Figure 5. It is observed that increased

Figure 4. Impact of (A) productivity and (B) lipid content on the

MFSP for a range of chemical coproduct prices.

broader productivity curve. The analysis highlights the

asymptotic cost responses, where productivity dramatically Figure 5. Impact of the split fraction of whole hydrolysate for chemical
inuences the MFSP below a value of roughly 0.5 g/L/hr and coproduction on the minimum fuel selling price for a range of
chemical coproduct prices.
then begins to approach a limit beyond which costs do not
signicantly improve (roughly 1 g/L/hr, which represents the
point at which bioconversion costs no longer dominate the production of the chemical coproduct relative to the fuel
overall system economics). If lipid productivity could be signicantly reduces the cost of the process. When 10%20%
improved to a hypothetical 2 g/L/hr, which is more of the hydrolysate stream is upgraded to chemicals, the
commensurate with the chemicals production step, the MFSP coproduct price has a minor impact on the MFSP (e.g., the
would decrease to $4.07/GGE. dierence is $0.55/GGE for the lowest and highest value
In the case of lipid content, if the lipid content in the cells coproduct when 10% of the hydrolysate is split to chemicals
could theoretically be improved from 70 to 95 wt % (although production). As the fraction of the hydrolysate upgraded to
such a high value is unlikely practical), the MFSP would chemicals increases beyond 30%, then the relative production
decrease by 10%. This suggests that for such large improve- split and the price of the coproduct have a greater impact on
the MFSP of the process.

ments in the process, the productivity reduces the MFSP by a
greater amount than the lipid content. However, if the focus
were on incremental improvements for near-term success, then DISCUSSION
a 25% improvement in lipid content would achieve a larger As demonstrated here, the inclusion of coproduct opportunities
reduction in the MFSP than a 25% improvement in into an integrated process design shows great potential to lower
productivity. Specically, a 25% enhancement in the lipid the MFSP of hydrocarbon biofuels produced from lignocellu-
content would reduce the MFSP by $0.38/GGE, while the losic biomass. The target case that includes production of a
same improvement in productivity would reduce the MFSP by chemical coproduct enables the potential to achieve an MFSP
$0.30/GGE. more than $4/GGE (45%) lower than the equivalent design
J DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

that solely produces fuel when maintaining the same perform- Consistent with established approaches for process design
ance and yield targets for fuel production. Given this key point, and engineering, the overall economics of a process can be
and in light of signicant cost sensitivities around the projected enhanced by understanding the extent of process integration
improved case performance parameters presented above, it is and identifying key approaches for intensication. For the
important to understand current experimental performance and biological conversion to fuels and chemicals considered here,
to prioritize research and development directions. When there are a myriad potential improvements. One of the primary
considering the current benchmark case for production of conclusions of our analysis is that for near-term incremental
fuels and chemicals, the MFSP is higher than either the targeted improvements the lipid content is a bigger cost driver than
fuel-only design or the fuels/chemicals design. Namely, the productivity for the process as long as productivity can remain
estimated benchmark MFSP is $11.37/GGE for the fuel/ over at least 0.3 g/L/hr. This may help guide R&D priorities, as
chemicals coproduction pathway, more than twice the it is well established that productivity and lipid content are
projected cost for the fuels/chemicals coproduction improved inversely related, i.e., operational strategies may be employed to
case and 20% higher than the fuel-only improved case. Figure target increased organism lipid content but typically at the
6 illustrates the variation of the cost breakout by process expense of slower production rates.
Another demonstrated advantage in the proposed design is
that the organisms being pursued are capable of producing
value-added products from a wide range of sugars and can
tolerate impurities common in hydrolysates. To date, promising
results have been reported in the literature for several
oleaginous yeast species on biomass-derived hydrolysates,
although substrate toxicity is still a major issue that limits
lipid content and productivity.70,89100 There are a similar
number of opportunities on the C6 train that involve
enhancing organism performance; for example, Blazeck et al.
describe an engineered Y. lipolytica strain that has been
observed to produce up to 71% lipids at a reasonable
productivity rate of 0.2 g/L/hr in small-scale batch culture
conditions.69 Higher lipid levels approaching 90% have also
been observed with variants of Y. lipolytica. 31 Quite
interestingly, Qiao et al. recently reported another engineered
variant of Y. lipolytica that is able to achieve 84.7% of theoretical
lipid yield at 55 g/L titers and a productivity of 0.71 g/L/hr.101
However, the bulk of these studies has been focused on small
scale cultivations with glucose, and such results will likely not to
directly translate to hydrolysates. Thus, while 90% lipids may be
too optimistic today for a realistic commercial-scale system
utilizing biomass hydrolysate, even incremental changes to both
the lipid content and the productivities would improve the
overall economics.
Beyond near-term organism targets based on intracellular
lipid pathways, there are other promising options with the
potential to signicantly improve process economics. For
example, published literature describes an autolysing yeast
strain currently utilized in the wine industry for its resulting
Figure 6. Cost breakout comparison of benchmark and improved
avor prole, whereby the cell is triggered to lyse and release its
cases for coproduction of fuels/chemicals scenario and improved case
for fuel-only scenario. The gray bar and reported total value intracellular contents without the need for a dedicated
correspond to the MFSP for each case. extraction step.102,103 If such an organism could be utilized
and/or engineered in this context for lipid accumulation and
self-induced release, the resulting cost would drop substantially
section for all three cases. Comparing the two improved to $4.18/GGE as shown previously in Figure 3. Furthermore,
cases, the overall cost breakouts are similar for all sections on a other organisms capable of product secretion (whether bacterial
$/GGE basis (in fact somewhat higher for the fuel/chemical or otherwise) possess the potential to reduce costs even further.
pathway given a lower fuel yield which increases $/GGE cost Cost savings brought about by removal of extraction capital and
contributions). It is clear that the reduction in the overall operating costs, combined with improved carbon eciencies to
MFSP cost is due to the chemical coproduct, which provides fuel, suggest signicant room for improved economics moving
for substantial coproduct revenue. When comparing the forward. Additionally, we stress that lipid production in
benchmark and improved cases for the coproduction of fuels oleaginous microbes are merely one of many variations of
and chemicals pathway, given the higher cost of the benchmark biological fuel precursor production, and indeed, many
case, a number of process improvements are needed moving strategies are being pursued that target other intermediates.
forward to reduce the overall cost of the design to the projected As demonstrated here, detailed TEA in each case will be of
values. It is promising, however, that there are a number of signicant importance to understand the technology drivers for
additional opportunities to further reduce costs. attaining cost-eective production of any given fuel precursor.
K DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

For the production of chemicals, A. succinogenes is able to integrated bioref inery, specically using our research data that
tolerate impurities common in hydrolysates, including furfural demonstrates biological production of succinic acid at
and HMF at concentrations relevant to biomass hydrolysate encouraging performance levels. Moreover, when viewed as
without substantial decreases in yield.84,104 It readily converts an intermediate, there is a strong potential for a greater demand
mixed sugar streams of both C6 and C5 sugars common in of succinic acid given that there are numerous known
hydrolysates. The capability of the organism to utilize minor technologies for the production of a wide slate of high-value
sugars, including arabinose and galactose, as well as to enhance large market commodity chemicals from succinic acid as
the yield for production when CO2 is included in the illustrated in Figure 7. On the basis of these potential routes to
conversion, may lead to improved carbon eciency in the chemicals alone, the market size for succinic acid has the
overall process and allow for an additional low cost feed stream potential to be well over 2,000,000 tons/year.60,110
and carbon source to further improve the economics of the
process. If the productivity of this organism could reach the 4
g/L/hr values reported for engineered E. coli105 on clean
commodity C6 sugars, the MFSP would decrease by $0.10/
GGE. It is important to further note that the overall succinic
acid yields from these E. coli strains are lower than those
demonstrated by A. succinogenes.
A major driver in the economics of the C5 chemicals
production train is the cost of separating succinic acid from the
fermentation broth. The bulk of this cost is not in purication
or polishing of the nal product but rather the requirement for
handling the succinate salt produced in the biological
conversion step. The need to maintain the pH of the
fermentation near neutral conditions dramatically increases
the complexity and operations for this separation. If the cost
associated with only the caustic added to the biological Figure 7. Known succinic acid derivatives.41,48,53,114,115
conversion could be completely removed, the MFSP would
decrease by over $1/GGE. In part, this could be accomplished The concept of improving the overall economics of a
by developing separation strategies in which the salt is biorenery through the production of high-value chemical
recovered and recycled.106,107 Another more promising coproducts from a fraction of lignocellulosic hydrolysate can be
approach would be to perform the biological upgrading at applied to a wide array of other intermediates.48,53,109,111113
low pH such that the addition of salt was not required, and the This modular approach will allow for exibility when
separation strategy could be simplied to completely eliminate developing bioreneries to avoid overwhelming a single
the need for the SMB unit. Approaches such as this are under individual commodity chemical market, while supporting the
development in acid-tolerant yeast such as Issatchenkia nancial feasibility using realistic process and market scenarios
orientalis.108 If this design strategy is possible and the yield of for modeling coproduct pathways. As outlined previously, the
the succinic acid remained constant, the overall MFSP would underlying price of the coproduct can have a large impact on
be substantially reduced to $2.32/GGE. If a low pH the MFSP as some products will not garner a high enough value
fermentation would not exhibit as high of a yield as that to overcome the additional cost in both capital and operating
being projected in this design, however, we estimate that the expenses attributed to additional processing steps.
succinic acid yield could decrease by about 18% and still Beyond the projections and additional improvements specic
achieve cost parity with the improved coproduction of fuels to fuels and chemicals production discussed here, additional
and chemicals case. Due to this clear potential cost advantage as opportunities exist regarding overall process integration to
well as the potential for a more simplied recovery scheme, a maximize yields to the desired products. The fuels and chemical
number of commercial companies producing organic acids have production strategies outlined can convert both C5 and C6
highlighted ongoing R&D to develop low pH fermentation sugars with a high tolerance to impurities in hydrolysates.
routes. Lastly, we also note that the details of the fermentation Customizing the biomass deconstruction strategies to maximize
and separations technology modeled here are specic to the desirable sugar intermediates through improvements to
succinic acid production; however, many other products can be pretreatment9,116 and cellulase eciency117 may further
produced biologically or catalytically from C5-enriched hydro- improve economics, while ensuring high eciencies of the
lysate. solidliquid separation strategies. Additionally, lignin, acetate,
It is also important to consider the rationale for coproduct and other components in biomass represent an opportunity for
additional coproducts.118122

selection and the implications of scale-up in terms of the
market and commercial biorenery economics (here specically
considering succinic acid). In 2009, worldwide consumption of CONCLUSION
succinic acid was 30,000 tons per year with a projected growth The detailed analysis of an integrated biorenery performed
of 5% per year.109 The current design would almost quadruple here presents (a) an economic potential for a whole
the 2009 production level with an estimated production rate of hydrolysate to hydrocarbon fuels process based on plausible
127,000 tons of succinic acid per year from a single biorenery near-term improvements to key conversion parameters and (b)
if succinic acid were intended to be the nal and only product. a similar potential for introducing a chemical coproduction
However, we stress that the intent for this design strategy is to pathway into the biorenery schematic. We show that the
utilize succinic acid as a representative molecule to demonstrate the inclusion of coproducts can dramatically improve the overall
potential for coproduct pathways within the f ramework of an economic viability of an integrated process for fuel production,
L DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

similar to the approach commonly employed in petroleum (8) Wyman, C. E.; Dale, B. E.; Elander, R. T.; Holtzapple, M.;
rening. However, there are underlying challenges as the Ladisch, M. R.; Lee, Y. Coordinated Development of Leading Biomass
additional processing steps increase the complexity of the Pretreatment Technologies. Bioresour. Technol. 2005, 96 (18), 1959
process and result in higher overall capital and operating costs 1966.
(9) Mosier, N.; Wyman, C.; Dale, B.; Elander, R.; Lee, Y.;
for this integrated design. The overall coproduct value also can
Holtzapple, M.; Ladisch, M. Features of Promising Technologies for
play a major role in the economics of the process. Key drivers Pretreatment of Lignocellulosic Biomass. Bioresour. Technol. 2005, 96
in the integrated design are tied to achievable yields within the (6), 673686.
individual biological conversion strategies and product (10) Lynd, L. R.; Van Zyl, W. H.; McBride, J. E.; Laser, M.
separations and also to improving the eciency in the Consolidated Bioprocessing of Cellulosic Biomass: An Update. Curr.
production and retention of sugars.

Opin. Biotechnol. 2005, 16 (5), 577583.

(11) O hgren, K.; Bura, R.; Lesnicki, G.; Saddler, J.; Zacchi, G. A
ASSOCIATED CONTENT Comparison between Simultaneous Saccharification and Fermentation
S Supporting Information and Separate Hydrolysis and Fermentation Using Steam-Pretreated
The Supporting Information is available free of charge on the Corn Stover. Process Biochem. 2007, 42 (5), 834839.
(12) Sheehan, J.; Aden, A.; Paustian, K.; Killian, K.; Brenner, J.;
ACS Publications website at DOI: 10.1021/acssusche-
Walsh, M.; Nelson, R. Energy and Environmental Aspects of Using
meng.6b00243. Corn Stover for Fuel Ethanol. J. Ind. Ecol. 2003, 7 (34), 117146.
Model overview, reaction stoichiometries, lipid recovery (13) Aden, A.; Foust, T. Technoeconomic Analysis of the Dilute
yields, and capital cost breakdown. (PDF) Sulfuric Acid and Enzymatic Hydrolysis Process for the Conversion of

Corn Stover to Ethanol. Cellulose 2009, 16 (4), 535545.

(14) Kazi, F. K.; Fortman, J. A.; Anex, R. P.; Hsu, D. D.; Aden, A.;
Dutta, A.; Kothandaraman, G. Techno-Economic Comparison of
Corresponding Authors Process Technologies for Biochemical Ethanol Production from Corn
*E-mail: (M.J.B.). Stover. Fuel 2010, 89, S20S28.
*E-mail: (G.T.B.). (15) Williams, P. R.; Inman, D.; Aden, A.; Heath, G. A.
Notes Environmental and Sustainability Factors Associated with Next-
The authors declare no competing nancial interest. Generation Biofuels in the Us: What Do We Really Know? Environ.

Sci. Technol. 2009, 43 (13), 47634775.

(16) Wooley, R.; Ruth, M.; Sheehan, J.; Ibsen, K.; Majdeski, H.;
Galvez, A. Lignocellulosic Biomass to Ethanol Process Design and
We thank the U.S. Department of Energy BioEnergy Economics Utilizing Co-Current Dilute Acid Prehydrolysis and Enzymatic
Technologies Oce for funding this work through Contract Hydrolysis Current and Futuristic Scenarios; NREL/TP-580-26157;
No. DE-AC36-08GO28308 at the National Renewable Energy National Renewable Energy Laboratory: Golden, CO, 1999.
Laboratory. We thank Rick Elander for a critical reading of the (17) Aden, A.; Ruth, M.; Ibsen, K.; Jechura, J.; Neeves, K.; Sheehan,
manuscript and Michael Bradeld, Adam Bratis, Rick Elander, J.; Wallace, B.; Montague, L.; Slayton, A.; Lukas, J. Lignocellulosic
Jacob Kruger, Jim McMillan, and Willie Nicol for helpful Biomass to Ethanol Process Design and Economics Utilizing Co-Current
discussions. The U.S. Government retains and the publisher, by Dilute Acid Prehydrolysis and Enzymatic Hydrolysis for Corn Stover;
NREL/TP -510-32438; National Renewable Energy Laboratory:
accepting the article for publication, acknowledges that the U.S.
Golden, CO, 2002.
Government retains a nonexclusive, paid up, irrevocable, (18) Hamelinck, C. N.; Hooijdonk, G. v.; Faaij, A. P. Ethanol from
worldwide license to publish or reproduce the published form Lignocellulosic Biomass: Techno-Economic Performance in Short-,
of this work, or allow others to do so, for U.S. Government Middle-and Long-Term. Biomass Bioenergy 2005, 28 (4), 384410.

(19) Klein-Marcuschamer, D.; Simmons, B. A.; Blanch, H. W.

Techno-Economic Analysis of a Lignocellulosic Ethanol Biorefinery
REFERENCES with Ionic Liquid Pre-Treatment. Biofuels, Bioprod. Biorefin. 2011, 5
(1) Pacala, S.; Socolow, R. Stabilization Wedges: Solving the Climate (5), 562569.
Problem for the Next 50 Years with Current Technologies. Science (20) Wright, M. M.; Brown, R. C. Comparative Economics of
2004, 305 (5686), 968972. Biorefineries Based on the Biochemical and Thermochemical
(2) Ragauskas, A.; Williams, C.; Davison, B.; Britovsek, G.; Cairney, Platforms. Biofuels, Bioprod. Biorefin. 2007, 1 (1), 4956.
J.; Eckert, C.; Frederick, W.; Hallett, J.; Leak, D.; Liotta, C.; Mielenz, (21) Sassner, P.; Galbe, M.; Zacchi, G. Techno-Economic Evaluation
J.; Murphy, R.; Templer, R.; Tschaplinski, T. The Path Forward for of Bioethanol Production from Three Different Lignocellulosic
Biofuels and Biomaterials. Science 2006, 311, 484489. Materials. Biomass Bioenergy 2008, 32 (5), 422430.
(3) Souza, G. M.; Victoria, R. L.; Joly, C. A.; Verdade, L. M. Bioenergy (22) Humbird, D.; Davis, R.; Tao, L.; Kinchin, C.; Hsu, D.; Aden, A.;
& Sustainability: Bridging the Gaps; Scientic Committee on Problems Schoen, P.; Lukas, J.; Olthof, B.; Worley, M. Process Design and
of the Environment (SCOPE), 2015. Economics for Biochemical Conversion of Lignocellulosic Biomass to
(4) Lane, J. Biofuels Mandates around the World; Biofuels Digest, Ethanol; NREL/TP-5100-47764; National Renewable Energy Labo-
December 31, 2014. ratory: Golden, CO, 2011.
(5) Tilman, D.; Socolow, R.; Foley, J. A.; Hill, J.; Larson, E.; Lynd, L.; (23) Jacobson, J. J.; Searcy, E.; Caerty, K.; Dunn, J. B.; Johnson, M.;
Pacala, S.; Reilly, J.; Searchinger, T.; Somerville, C.; Williams, R. Wang, Z.; Wang, M.; Biddy, M.; Dutta, A.; Inman, D. Supply Chain
Beneficial Biofuelsthe Food, Energy, and Environment Trilemma. Sustainability Analysis of Three Biofuel Pathways; ANL/ESD-14/5;
Science 2009, 325 (5938), 270. Argonne National Laboratory (ANL): Argonne, IL, 2013.
(6) Himmel, M. E.; Ding, S.-Y.; Johnson, D. K.; Adney, W. S.; (24) Canter, C. E.; Dunn, J. B.; Han, J.; Wang, Z.; Wang, M. Policy
Nimlos, M. R.; Brady, J. W.; Foust, T. D. Biomass Recalcitrance: Implications of Allocation Methods in the Life Cycle Analysis of
Engineering Plants and Enzymes for Biofuels Production. Science 2007, Integrated Corn and Corn Stover Ethanol Production. BioEnergy Res.
315 (5813), 804807. 2016, 9 (1), 7787.
(7) Chundawat, S. P. S.; Beckham, G. T.; Himmel, M. E.; Dale, B. E. (25) Murphy, C. W.; Kendall, A. Life Cycle Analysis of Biochemical
Deconstruction of Lignocellulosic Biomass to Fuels and Chemicals. Cellulosic Ethanol under Multiple Scenarios. GCB Bioenergy 2015, 7
Annu. Rev. Chem. Biomol. Eng. 2011, 2, 121145. (5), 10191033.

M DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

(26) Peralta-Yahya, P. P.; Zhang, F.; del Cardayre, S. B.; Keasling, J. (45) Michinaka, Y.; Shimauchi, T.; Aki, T.; Nakajima, T.; Kawamoto,
D. Microbial Engineering for the Production of Advanced Biofuels. S.; Shigeta, S.; Suzuki, O.; Ono, K. Extracellular Secretion of Free Fatty
Nature 2012, 488 (7411), 320328. Acids by Disruption of a Fatty Acyl-Coa Synthetase Gene in
(27) Serrano-Ruiz, J. C.; Dumesic, J. A. Catalytic Routes for the Saccharomyces Cerevisiae. J. Biosci. Bioeng. 2003, 95 (5), 435440.
Conversion of Biomass into Liquid Hydrocarbon Transportation (46) Leber, C.; Polson, B.; Fernandez-Moya, R.; Da Silva, N. A.
Fuels. Energy Environ. Sci. 2011, 4 (1), 8399. Overproduction and Secretion of Free Fatty Acids through Disrupted
(28) Narula, C. K.; Li, Z.; Casbeer, E. M.; Geiger, R. A.; Moses- Neutral Lipid Recycle in Saccharomyces Cerevisiae. Metab. Eng. 2015,
Debusk, M.; Keller, M.; Buchanan, M. V.; Davison, B. H., 28, 5462.
Heterobimetallic Zeolite, Inv-Zsm-5, Enables Ecient Conversion of (47) Scharnewski, M.; Pongdontri, P.; Mora, G.; Hoppert, M.; Fulda,
Biomass Derived Ethanol to Renewable Hydrocarbons. Sci. Rep. 2015, M. Mutants of Saccharomyces cerevisiae Deficient in Acyl-Coa
5.1603910.1038/srep16039 Synthetases Secrete Fatty Acids Due to Interrupted Fatty Acid
(29) Mettler, M. S.; Vlachos, D. G.; Dauenhauer, P. J. Top Ten Recycling. FEBS J. 2008, 275 (11), 27652778.
Fundamental Challenges of Biomass Pyrolysis for Biofuels. Energy (48) Werpy, T.; Petersen, G.; Aden, A.; Bozell, J.; Holladay, J.; White,
Environ. Sci. 2012, 5 (7), 77977809. J.; Manheim, A.; Eliot, D.; Lasure, L.; Jones, S. Top Value Added
(30) Luque, R.; de la Osa, A. R.; Campelo, J. M.; Romero, A. A.; Chemicals from Biomass. Volume 1Results of Screening for Potential
Valverde, J. L.; Sanchez, P. Design and Development of Catalysts for Candidates from Sugars and Synthesis Gas; Pacic Northwest National
Biomass-to-Liquid-FischerTropsch (Btl-Ft) Processes for Biofuels Laboratory and National Renewable Energy Laboratory, 2004.
Production. Energy Environ. Sci. 2012, 5 (1), 51865202. (49) Corma, A.; Iborra, S.; Velty, A. Chemical Routes for the
(31) Van de Vyver, S.; Geboers, J.; Jacobs, P. A.; Sels, B. F. Recent Transformation of Biomass into Chemicals. Chem. Rev. 2007, 107 (6),
Advances in the Catalytic Conversion of Cellulose. ChemCatChem 24112502.
2011, 3 (1), 8294. (50) Bozell, J. J.; Holladay, J. E.; White, J. F.; Johnson, D. K. Top
(32) de Beeck, B. O.; Dusselier, M.; Geboers, J.; Holsbeek, J.; Morre, Value-Added Chemicals from Biomass. Volume IIResults of Screening for
E.; Oswald, S.; Giebeler, L.; Sels, B. F. Direct Catalytic Conversion of Potential Candidates from Biorenery Lignin; Pacic Northwest National
Cellulose to Liquid Straight-Chain Alkanes. Energy Environ. Sci. 2015, Laboratory, National Renewable Energy Laboratory, and University of
8, 230. Tennessee, 2007
(33) Prasomsri, T.; Nimmanwudipong, T.; Roman-Leshkov, Y. (51) Amidon, T. E.; Wood, C. D.; Shupe, A. M.; Wang, Y.; Graves,
Effective Hydrodeoxygenation of Biomass-Derived Oxygenates into M.; Liu, S. Biorefinery: Conversion of Woody Biomass to Chemicals,
Unsaturated Hydrocarbons by Moo 3 Using Low H 2 Pressures. Energy and Materials. J. Biobased Mater. Bioenergy 2008, 2 (2), 100
Energy Environ. Sci. 2013, 6 (6), 17321738. 120.
(34) Rinaldi, R.; Schuth, F. Design of Solid Catalysts for the
(52) Bozell, J. J. Feedstocks for the FutureBiorefinery Production of
Conversion of Biomass. Energy Environ. Sci. 2009, 2 (6), 610626.
Chemicals from Renewable Carbon. Clean: Soil, Air, Water 2008, 36
(35) Besson, M.; Gallezot, P.; Pinel, C. Conversion of Biomass into
(8), 641647.
Chemicals over Metal Catalysts. Chem. Rev. 2014, 114 (3), 1827
(53) Bozell, J. J.; Petersen, G. R. Technology Development for the
Production of Biobased Products from Biorefinery Carbohydrates-the
(36) Serrano-Ruiz, J. C.; Luque, R.; Sepulveda-Escribano, A.
Us Department of Energys Top 10 Revisited. Green Chem. 2010, 12
Transformations of Biomass-Derived Platform Molecules: From
High Added-Value Chemicals to Fuelsvia Aqueous-Phase Processing. (4), 539554.
(54) Vennestrm, P.; Osmundsen, C. M.; Christensen, C.; Taarning,
Chem. Soc. Rev. 2011, 40 (11), 52665281.
(37) Lange, J.-P. Renewable Feedstocks: The Problem of Catalyst E. Beyond Petrochemicals: The Renewable Chemicals Industry.
Deactivation and Its Mitigation. Angew. Chem., Int. Ed. 2015, 54 (45), Angew. Chem., Int. Ed. 2011, 50 (45), 1050210509.
1318613197. (55) Wettstein, S. G.; Alonso, D. M.; Gurbuz, E. I.; Dumesic, J. A. A
(38) Lee, S. K.; Chou, H.; Ham, T. S.; Lee, T. S.; Keasling, J. D. Roadmap for Conversion of Lignocellulosic Biomass to Chemicals and
Metabolic Engineering of Microorganisms for Biofuels Production: Fuels. Curr. Opin. Chem. Eng. 2012, 1 (3), 218224.
From Bugs to Synthetic Biology to Fuels. Curr. Opin. Biotechnol. 2008, (56) Gallezot, P. Conversion of Biomass to Selected Chemical
19 (6), 556563. Products. Chem. Soc. Rev. 2012, 41 (4), 15381558.
(39) Klinke, H. B.; Thomsen, A.; Ahring, B. K. Inhibition of Ethanol- (57) Tuck, C. O.; Perez, E.; Horvath, I. T.; Sheldon, R. A.; Poliakoff,
Producing Yeast and Bacteria by Degradation Products Produced M. Valorization of Biomass: Deriving More Value from Waste. Science
During Pre-Treatment of Biomass. Appl. Microbiol. Biotechnol. 2004, 2012, 337 (6095), 695699.
66 (1), 1026. (58) Koutinas, A. A.; Vlysidis, A.; Pleissner, D.; Kopsahelis, N.; Lopez
(40) Pienkos, P. T.; Zhang, M. Role of Pretreatment and Garcia, I.; Kookos, I. K.; Papanikolaou, S.; Kwan, T. H.; Lin, C. S. K.
Conditioning Processes on Toxicity of Lignocellulosic Biomass Valorization of Industrial Waste and by-Product Streams Via
Hydrolysates. Cellulose 2009, 16 (4), 743762. Fermentation for the Production of Chemicals and Biopolymers.
(41) Davis, R.; Tao, L.; Tan, E.; Biddy, M. J.; Beckham, G. T.; Chem. Soc. Rev. 2014, 43 (8), 25872627.
Scarlata, C.; Jacobson, J.; Caerty, K.; Ross, J.; Lukas, J.; Knorr, D.; (59) Dusselier, M.; Mascal, M.; Sels, B. Top Chemical Opportunities
Schoen, P. Process Design and Economics for the Conversion of from Carbohydrate Biomass: A Chemists View of the Biorenery. In
Lignocellulosic Biomass to Hydrocarbons: Dilute-Acid Prehydrolysis and Selective Catalysis for Renewable Feedstocks and Chemicals; Nicholas, K.
Enzymatic Hydrolysis Deconstruction of Biomass to Sugars and Biological M., Ed.; Springer International Publishing: 2014; Vol. 353, pp 140.
Conversion of Sugars to Hydrocarbons; NREL/TP-5100-60223; Na- (60) Cheng, K.-K.; Zhao, X.-B.; Zeng, J.; Zhang, J.-A. Biotechno-
tional Renewable Energy Laboratory: Golden, CO, 2013. logical Production of Succinic Acid: Current State and Perspectives.
(42) Chen, X.; Shekiro, J.; Franden, M. A.; Wang, W.; Zhang, M.; Biofuels, Bioprod. Biorefin. 2012, 6 (3), 302318.
Kuhn, E.; Johnson, D. K.; Tucker, M. P. The Impacts of Deacetylation (61) Tao, L.; Schell, D.; Davis, R.; Tan, E.; Elander, R. T.; Bratis, A.
Prior to Dilute Acid Pretreatment on the Bioethanol Process. NREL 2012 Achievement of Ethanol Cost Targets: Biochemical Ethanol
Biotechnol. Biofuels 2012, 5, 8. Fermentation via Dilute-Acid Pretreatment and Enzymatic Hydrolysis of
(43) Liu, H.; Yu, C.; Feng, D.; Cheng, T.; Meng, X.; Liu, W.; Zou, Corn Stover; NREL/TP-5100-61563; National Renewable Energy
H.; Xian, M. Production of Extracellular Fatty Acid Using Engineered Laboratory: Golden, CO, 2014.
Escherichia Coli. Microb. Cell Fact. 2012, 11 (1), 4154. (62) Sievers, D. A.; Tao, L.; Schell, D. J. Performance and Techno-
(44) Meng, X.; Shang, H.; Zheng, Y.; Zhang, Z. Free Fatty Acid Economic Assessment of Several SolidLiquid Separation Technol-
Secretion by an Engineered Strain of Escherichia Coli. Biotechnol. Lett. ogies for Processing Dilute-Acid Pretreated Corn Stover. Bioresour.
2013, 35 (12), 20992103. Technol. 2014, 167, 291296.

N DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

(63) Runguphan, W.; Keasling, J. D. Metabolic Engineering of (83) Liu, Y.-P.; Zheng, P.; Sun, Z.-H.; Ni, Y.; Dong, J.-J.; Zhu, L.-L.
Saccharomyces Cerevisiae for Production of Fatty Acid-Derived Economical Succinic Acid Production from Cane Molasses by
Biofuels and Chemicals. Metab. Eng. 2014, 21, 103113. Actinobacillus Succinogenes. Bioresour. Technol. 2008, 99 (6), 1736
(64) Davis, M. S.; Solbiati, J.; Cronan, J. E. Overproduction of Acetyl- 1742.
Coa Carboxylase Activity Increases the Rate of Fatty Acid Biosynthesis (84) Bradfield, M. F.; Mohagheghi, A.; Salvachua, D.; Smith, H.;
in Escherichia Coli. J. Biol. Chem. 2000, 275 (37), 2859328598. Black, B. A.; Dowe, N.; Beckham, G. T.; Nicol, W. Continuous
(65) Lu, X.; Vora, H.; Khosla, C. Overproduction of Free Fatty Acids Succinic Acid Production by Actinobacillus Succinogenes on Xylose-
in E. Coli: Implications for Biodiesel Production. Metab. Eng. 2008, 10 Enriched Hydrolysate. Biotechnol. Biofuels 2015, 8 (1), 1.
(6), 333339. (85) Gerberding, S. J.; Singh, R., Purication of Succinic Acid from
(66) Clomburg, J. M.; Gonzalez, R. Biofuel Production in Escherichia the Fermentation Broth Containing Ammonium Succinate. EP Patent
Coli: The Role of Metabolic Engineering and Synthetic Biology. Appl. EP2519491 A2, 2010.
Microbiol. Biotechnol. 2010, 86 (2), 419434. (86) Davis, R.; Tao, L.; Scarlata, C.; Tan, E. C. D.; Ross, J.; Lukas, J.;
(67) Beopoulos, A.; Cescut, J.; Haddouche, R.; Uribelarrea, J.-L.; Sexton, D. Process Design and Economics for the Conversion of
Molina-Jouve, C.; Nicaud, J.-M. Yarrowia Lipolytica as a Model for Lignocellulosic Biomass to Hydrocarbons: Dilute-Acid and Enzymatic
Bio-Oil Production. Prog. Lipid Res. 2009, 48 (6), 375387. Deconstruction of Biomass to Sugars and Catalytic Conversion of Sugars to
(68) Tai, M.; Stephanopoulos, G. Engineering the Push and Pull of Hydrocarbons; NREL/TP-5100-62498; National Renewable Energy
Lipid Biosynthesis in Oleaginous Yeast Yarrowia Lipolytica for Biofuel Laboratory: Golden, CO, 2015.
Production. Metab. Eng. 2013, 15, 19. (87) Tan, E. C. D.; Talmadge, M.; Dutta, A.; Hensley, J.; Schaidle, J.;
(69) Blazeck, J.; Hill, A.; Liu, L.; Knight, R.; Miller, J.; Pan, A.; Biddy, M.; Humbird, D.; Snowden-Swan, L. J.; Ross, J.; Sexton, D.;
Otoupal, P.; Alper, H. S. Harnessing Yarrowia Lipolytica Lipogenesis Yap, R.; Lukas, J. Process Design and Economics for the Conversion of
to Create a Platform for Lipid and Biofuel Production. Nat. Commun. Lignocellulosic Biomass to Hydrocarbons via Indirect Liquefaction:
2014, DOI: 10.1038/ncomms4131. Thermochemical Research Pathway to High-Octane Gasoline Blendstock
(70) Li, Y.; Zhao, Z. K.; Bai, F. High-Density Cultivation of through Methanol/Dimethyl Ether Intermediates; NREL/TP-5100-
Oleaginous Yeast Rhodosporidium Toruloides Y4 in Fed-Batch 62402; National Renewable Energy Laboratory: Golden, CO, 2015.
Culture. Enzyme Microb. Technol. 2007, 41 (3), 312317. (88) Jones, S. B.; Meyer, P. A.; Snowden-Swan, L. J.; Padmaperuma,
(71) Zhu, Z.; Zhang, S.; Liu, H.; Shen, H.; Lin, X.; Yang, F.; Zhou, Y. A. B.; Tan, E. C. D.; Dutta, A.; Jacobson, J.; Caerty, K. Process Design
J.; Jin, G.; Ye, M.; Zou, H.; Zhao, Z. K. A Multi-Omic Map of the and Economics for the Conversion of Lignocellulosic Biomass to
Lipid-Producing Yeast Rhodosporidium Toruloides. Nat. Commun. Hydrocarbon Fuels: Fast Pyrolysis and Hydrotreating Bio-Oil Pathway;
2012, 3, 1112. NREL/TP-5100-61178; Pacic Northwest National Laboratory,
(72) Zhao, X.; Kong, X.; Hua, Y.; Feng, B.; Zhao, Z. K. Medium National Renewable Energy Laboratory, and Idaho National
Optimization for Lipid Production through Co-Fermentation of Laboratory: 2013.
(89) Galafassi, S.; Cucchetti, D.; Pizza, F.; Franzosi, G.; Bianchi, D.;
Glucose and Xylose by the Oleaginous Yeast Lipomyces Starkeyi.
Compagno, C. Lipid Production for Second Generation Biodiesel by
Eur. J. Lipid Sci. Technol. 2008, 110 (5), 405412.
the Oleaginous Yeast Rhodotorula Graminis. Bioresour. Technol. 2012,
(73) Gong, Z.; Wang, Q.; Shen, H.; Hu, C.; Jin, G.; Zhao, Z. K. Co-
111, 398403.
Fermentation of Cellobiose and Xylose by Lipomyces Starkeyi for
(90) Huang, C.; Chen, X.-f.; Xiong, L.; Yang, X.-y.; Chen, X.-d.; Ma,
Lipid Production. Bioresour. Technol. 2012, 117, 2024.
L.-l.; Chen, Y. Microbial Oil Production from Corncob Acid
(74) Huang, C.; Chen, X.-F.; Yang, X.-Y.; Xiong, L.; Lin, X.-Q.; Yang,
Hydrolysate by Oleaginous Yeast Trichosporon Coremiiforme.
J.; Wang, B.; Chen, X.-D. Bioconversion of Corncob Acid Hydrolysate
Biomass Bioenergy 2013, 49, 273278.
into Microbial Oil by the Oleaginous Yeast Lipomyces Starkeyi. Appl. (91) Huang, C.; Wu, H.; Li, R.-f.; Zong, M.-h. Improving Lipid
Biochem. Biotechnol. 2014, 172 (4), 21972204. Production from Bagasse Hydrolysate with Trichosporon Fermentans
(75) Laurens, L. M. L.; Nagle, N.; Davis, R.; Sweeney, N.; Van by Response Surface Methodology. New Biotechnol. 2012, 29 (3),
Wychen, S.; Lowell, A.; Pienkos, P. T. Acid-Catalyzed Algal Biomass 372378.
Pretreatment for Integrated Lipid and Carbohydrate-Based Biofuels (92) Chang, Y.-H.; Chang, K.-S.; Hsu, C.-L.; Chuang, L.-T.; Chen,
Production. Green Chem. 2015, 17 (2), 11451158. C.-Y.; Huang, F.-Y.; Jang, H.-D. A Comparative Study on Batch and
(76) Dong, T.; Knoshaug, E.; Davis, R.; Laurens, L.; Wychen, S. V.; Fed-Batch Cultures of Oleaginous Yeast Cryptococcus Sp. In Glucose-
Pienkos, P. T.; Nagle, N. Combined Algal Processing: A Novel Based Media and Corncob Hydrolysate for Microbial Oil Production.
Integrated Biorefinery Process to Produce Algal Biofuels and Fuel 2013, 105, 711717.
Bioproducts. Algal Res. 2016, DOI: 10.1016/j.algal.2015.12.021. (93) Huang, C.; Chen, X.-f.; Xiong, L.; Chen, X.-d.; Ma, L.-l. Oil
(77) Marker, T. L.; Petri, J.; Kalnes, T.; McCall, M.; Mackowiak, D.; Production by the Yeast Trichosporon Dermatis Cultured in
Jerosky, B.; Reagan, B.; Nemeth, L.; Krawyczyk, M.; Czernik, S.; Enzymatic Hydrolysates of Corncobs. Bioresour. Technol. 2012, 110,
Elliott, D.; Shonnard, D. Opportunities for Biorenewables in Oil 711714.
Reneries; Final Report; UOP: Des Plaines, IL, 2005. (94) Tsigie, Y. A.; Wang, C.-Y.; Truong, C.-T.; Ju, Y.-H. Lipid
(78) Dillich, S.; Ramsden, T.; Melina, M. U.S. Department of Energy Production from Yarrowia Lipolytica Po1g Grown in Sugarcane
Hydrogen and Fuel Cells Program, Program Records; U.S. Department of Bagasse Hydrolysate. Bioresour. Technol. 2011, 102 (19), 92169222.
Energy: 2012. (95) Yu, X.; Zeng, J.; Zheng, Y.; Chen, S. Effect of Lignocellulose
(79) Sievers, D. A.; Lischeske, J. J.; Biddy, M. J.; Stickel, J. J. A Low- Degradation Products on Microbial Biomass and Lipid Production by
Cost SolidLiquid Separation Process for Enzymatically Hydrolyzed the Oleaginous Yeast Cryptococcus Curvatus. Process Biochem. 2014,
Corn Stover Slurries. Bioresour. Technol. 2015, 187, 3742. 49 (3), 457465.
(80) Guettler, M. V.; Rumler, D.; Jain, M. K. Actinobacillus (96) Yu, X.; Zheng, Y.; Dorgan, K. M.; Chen, S. Oil Production by
Succinogenes Sp. Nov., a Novel Succinic-Acid-Producing Strain from Oleaginous Yeasts Using the Hydrolysate from Pretreatment of Wheat
the Bovine Rumen. Int. J. Syst. Bacteriol. 1999, 49 (1), 207216. Straw with Dilute Sulfuric Acid. Bioresour. Technol. 2011, 102 (10),
(81) Van der Werf, M. J.; Guettler, M. V.; Jain, M. K.; Zeikus, J. G. 61346140.
Environmental and Physiological Factors Affecting the Succinate (97) Zeng, J.; Zheng, Y.; Yu, X.; Yu, L.; Gao, D.; Chen, S.
Product Ratio During Carbohydrate Fermentation by Actinobacillus Lignocellulosic Biomass as a Carbohydrate Source for Lipid
Sp. 130z. Arch. Microbiol. 1997, 167 (6), 332342. Production by Mortierella Isabellina. Bioresour. Technol. 2013, 128,
(82) Yu, J.; Li, Z.; Ye, Q.; Yang, Y.; Chen, S. Development of Succinic 385391.
Acid Production from Corncob Hydrolysate by Actinobacillus (98) Yousuf, A. Biodiesel from Lignocellulosic Biomass Prospects
Succinogenes. J. Ind. Microbiol. Biotechnol. 2010, 37 (10), 10331040. and Challenges. Waste Manage. 2012, 32 (11), 20612067.

O DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX
ACS Sustainable Chemistry & Engineering Research Article

(99) Xue, Y.-P.; Jin, M.; Orjuela, A.; Slininger, P. J.; Dien, B. S.; Dale, M.; Langan, P.; Naskar, A. K.; Saddler, J. N.; Tschaplinski, T. J.;
B. E.; Balan, V. Microbial Lipid Production from Afex[Trade Mark Tuskan, G. A.; Wyman, C. E. Lignin Valorization: Improving Lignin
Sign] Pretreated Corn Stover. RSC Adv. 2015, 5 (36), 2872528734. Processing in the Biorefinery. Science 2014, 344 (6185), 1246843.
(100) Jin, M.; Slininger, P. J.; Dien, B. S.; Waghmode, S.; Moser, B. (119) Linger, J. G.; Vardon, D. R.; Guarnieri, M. T.; Karp, E. M.;
R.; Orjuela, A.; Sousa, L. d. C.; Balan, V. Microbial Lipid-Based Hunsinger, G. B.; Franden, M. A.; Johnson, C. W.; Chupka, G.;
Lignocellulosic Biorefinery: Feasibility and Challenges. Trends Strathmann, T. J.; Pienkos, P. T.; Beckham, G. T. Lignin Valorization
Biotechnol. 2015, 33 (1), 4354. through Integrated Biological Funneling and Chemical Catalysis. Proc.
(101) Qiao, K.; Imam Abidi, S. H.; Liu, H.; Zhang, H.; Chakraborty, Natl. Acad. Sci. U. S. A. 2014, 111 (33), 1201312018.
S.; Watson, N.; Kumaran Ajikumar, P.; Stephanopoulos, G. Engineer- (120) Vardon, D. R.; Franden, M. A.; Johnson, C. W.; Karp, E. M.;
ing Lipid Overproduction in the Oleaginous Yeast Yarrowia Lipolytica. Guarnieri, M. T.; Linger, J. G.; Salm, M. J.; Strathmann, T. J.;
Metab. Eng. 2015, 29, 5665. Beckham, G. T. Adipic Acid Production from Lignin. Energy Environ.
(102) Tabera, L.; Munoz, R.; Gonzalez, R. Deletion of Bcy1 from the Sci. 2015, 8 (2), 617.
Saccharomyces Cerevisiae Genome Is Semidominant and Induces (121) Zakzeski, J.; Bruijnincx, P. C. A.; Jongerius, A. L.; Weckhuysen,
Autolytic Phenotypes Suitable for Improvement of Sparkling Wines. B. M. The Catalytic Valorization of Lignin for the Production of
Appl. Environ. Microbiol. 2006, 72 (4), 23512358. Renewable Chemicals. Chem. Rev. 2010, 110 (6), 35523599.
(103) Cebollero, E.; Martinez-Rodriguez, A.; Carrascosa, A. V.; (122) Salvachua, D.; Karp, E. M.; Nimlos, C. T.; Vardon, D. R.;
Gonzalez, R. Overexpression of Csc11. A Plausible Strategy to Beckham, G. T. Towards Lignin Consolidated Bioprocessing:
Obtain Wine Yeast Strains Undergoing Accelerated Autolysis. FEMS Simultaneous Lignin Depolymerization and Product Generation by
Microbiol. Lett. 2005, 246 (1), 19. Bacteria. Green Chem. 2015, 17 (11), 49514967.
(104) Salvachua, D.; Mohagheghi, A.; Smith, H.; Bradfield, M. F. A.;
Nicol, W.; Black, B. A.; Biddy, M. J.; Dowe, N.; Beckham, G. T.
Succinic Acid Production on Xylose-Enriched Biorefinery Streams by
Actinobacillus Succinogenes in Batch Fermentation. Biotechnol. Biofuels
2016, DOI: 10.1186/s13068-016-0425-1.
(105) Jantama, K.; Haupt, M. J.; Zhang, X.; Moore, J. C.;
Shanmugam, K. T.; Ingram, L. O. N. Materials and Methods for
Ecient Succinate and Malate Production. World Patent
WO2008115958A3, 2014.
(106) Urbanus, J.; Roelands, C. P. M.; Verdoes, D.; ter Horst, J. H.
Intensified Crystallization in Complex Media: Heuristics for
Crystallization of Platform Chemicals. Chem. Eng. Sci. 2012, 77, 18
(107) Datta, R.; Glassner, D. A.; Jain, M. K.; Roy, J. R. V.
Fermentation and Purication Process for Succinic Acid. U.S. Patent
5168055, 1992.
(108) Xiao, H.; Shao, Z.; Jiang, Y.; Dole, S.; Zhao, H. Exploiting
Issatchenkia Orientalis Sd108 for Succinic Acid Production. Microb.
Cell Fact. 2014, DOI: 10.1186/s12934-014-0121-4.
(109) de Jong, E.; Higson, A.; Walsh, P.; Wellisch, M. Product
Developments in the Bio-Based Chemicals Arena. Biofuels, Bioprod.
Biorefin. 2012, 6 (6), 606624.
(110) Guzman, D. D. Bio-Succinic Acid Market Ready to Roar. ICIS
Chemical Business 2012, 281 (5), 28.
(111) Sheldon, R. A. Green and Sustainable Manufacture of
Chemicals from Biomass: State of the Art. Green Chem. 2014, 16
(3), 950963.
(112) Yim, H.; Haselbeck, R.; Niu, W.; Pujol-Baxley, C.; Burgard, A.;
Boldt, J.; Khandurina, J.; Trawick, J. D.; Osterhout, R. E.; Stephen, R.;
et al. Metabolic Engineering of Escherichia Coli for Direct Production
of 1,4-Butanediol. Nat. Chem. Biol. 2011, 7 (7), 445452.
(113) Dusselier, M.; Van Wouwe, P.; Dewaele, A.; Makshina, E.; Sels,
B. F. Lactic Acid as a Platform Chemical in the Biobased Economy:
The Role of Chemocatalysis. Energy Environ. Sci. 2013, 6 (5), 1415
(114) Dietrich, J. US Maleic Anhydride Contracts Flat. ICIS Chemical
Business 2014, 285 (25), 2121.
(115) Ebert, J. The Quest to Commercialize Biobased Succinic Acid.
Biomass Magazine, 2007
(116) Chen, X.; Wang, W.; Ciesielski, P. N.; Trass, O.; Park, S.; Tao,
L.; Tucker, M. Improving Sugar Yields and Reducing Enzyme
Loadings in the Deacetylation and Mechanical Refining (DMR)
Process through Multi-Stage Disk and Szego Refining and
Corresponding Techno Economic Analysis. ACS Sustainable Chem.
Eng. 2016, 4, 324.
(117) Payne, C. M.; Knott, B. C.; Mayes, H. B.; Hansson, H.;
Himmel, M. E.; Sandgren, M.; Stahlberg, J.; Beckham, G. T. Fungal
Cellulases. Chem. Rev. 2015, 115 (3), 13081448.
(118) Ragauskas, A. J.; Beckham, G. T.; Biddy, M. J.; Chandra, R.;
Chen, F.; Davis, M. F.; Davison, B. H.; Dixon, R. A.; Gilna, P.; Keller,

P DOI: 10.1021/acssuschemeng.6b00243
ACS Sustainable Chem. Eng. XXXX, XXX, XXXXXX