Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech
Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan
Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan
h i g h l i g h t s
" Biorenery approaches produce fuels and chemicals through biomass conversion.
" Consolidated bioprocessing of lignocellulose is desired for effective biorenery.
" This review focuses on the development of microbes for consolidated bioprocessing.
" The production of bio-based chemicals and advances fuels is emphasized.
a r t i c l e
i n f o
Article history:
Available online 23 October 2012
Keywords:
Biomass
Biorenery
Cellulase
Consolidated bioprocessing
Lignocellulose
a b s t r a c t
The biorenery manufacturing process for producing chemicals and liquid fuels from biomass is a promising approach for securing energy and resources. To establish cost-effective fermentation of lignocellulosic biomass, the consolidation of sacccharication and fermentation processes is a desirable strategy,
but requires the development of microorganisms capable of cellulose/hemicellulose hydrolysis and target
chemical production. Such an endeavor requires a large number of prerequisites to be realized, including
engineering microbial strains with high cellulolytic activity, high product yield, productivities, and titers,
ability to use many carbon sources, and resistance to toxic compounds released during the pretreatment
of lignocellulosic biomass. Researchers have focused on either engineering naturally cellulolytic microorganisms to improve product-related properties or modifying non-cellulolytic organisms with high product yields to become cellulolytic. This article reviews recent advances in the development of
microorganisms for the production of renewable chemicals and advanced biofuels, as well as ethanol,
from lignocellulosic materials through consolidated bioprocessing.
2012 Elsevier Ltd. All rights reserved.
1. Introduction
To ensure a reliable future source of energy and raw materials,
the utilization of sustainable biomass has considerable advantages
over petroleum-based energy sources. A biorenery is a concept
that integrates biomass conversion processes and equipment to
produce fuels, power, and chemicals from biomass, instead of conventional oil renery processes. Lignocellulosic biomass obtained
as agriculture byproducts and industrial residues is an abundant,
inexpensive, and renewable source of sugars, and is a desirable
feedstock for the sustainable production of liquid fuels and chemical products through the biorenery processes (Menon and Rao,
2012). After pretreatment of the biomass, cellulosic and hemicellulosic materials are enzymatically decomposed into simple sugars
that can be metabolized by microorganisms and converted to
Corresponding author. Tel./fax: +81 78 803 6196.
E-mail address: akondo@kobe-u.ac.jp (A. Kondo).
0960-8524/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.biortech.2012.10.047
514
2012). However, the hydrolysis of cellulose remains a major limiting factor for the efcient utilization of lignocellulosic materials
(Matano et al., 2012; Olson et al., 2011).
To release soluble sugars from cellulose, the activities of multiple enzymes, including endoglucanase, exoglucanase, and b-glucosidase, are required (Chandel et al., 2012). As cellulase reactions are
inhibited by their intermediary and nal products, such as cellooligosaccharides and glucose, microbial fermentation processes that
combine enzymatic hydrolysis with sugar consumption are preferential for the alleviation of cellulase activity inhibition (van Zyl
et al., 2007). However, the large difference in optimum temperatures between saccharication and fermentation during simultaneous saccharication and fermentation (SSF) is a drawback of
bio-ethanol production (Hasunuma and Kondo, 2012). To overcome this limitation, large amounts of saccharication enzymes
by fungi and bacteria are required, which severely impacts the cost
effectiveness of biorenery from lignocellulosic materials.
The recent development of microorganisms capable of efcient
cellulose hydrolysis and fermentation represents a signicant step
in reducing the requirement for enzyme addition into the SSF processes (Matano et al., 2012). Such consolidation of enzyme production, saccharication, and fermentation into a single process is
increasingly recognized as having potential for the low-cost production of biofuels and bio-based chemicals, as the high costs of
capital investment, raw materials, and equipment associated with
microbial enzyme production can be avoided (Menon and Rao,
2012; Mussatto et al., 2010; Olson et al., 2011). To develop such
organisms, researchers have focused on either engineering naturally cellulolytic microorganisms to improve product-related properties or modifying non-cellulolytic organisms with high product
yields to become cellulolytic (Hasunuma and Kondo, 2011; Olson
et al., 2011), as there is still no ideal organism to use in one-step
biomass conversion.
Although early efforts toward achieving efcient biorenery
processes from plant biomass have typically focused on ethanol,
recent advances in microbial metabolic engineering have enabled
to perform a challenge for producing renewable chemicals and advanced bio-fuels that are compatible with existing engines and fuel
distribution infrastructure (Zhang et al., 2011a). This review will
focus on a discussion of microbial strains for use in consolidated
bioprocessing (CBP) technologies. In particular, approaches for
the conversion of lignocellulosic materials into bio-based chemicals and fuels, as well as bio-ethanol, through microbial fermentation are emphasized.
515
Francisco et al. (1993) used an immobilization technique to prepare whole-cell biocatalysts of an E. coli strain that specically
binds to cellulosic materials. Genes of the Cellulomonas mi exoglucanase, Cex, and its cellulose-binding domain, CBDCex, were expressed in E. coli as fusion proteins with Lpp-OmpA, consisting of
the major outer membrane lipoprotein, Lpp, and amino acids
46159 of the outer membrane protein, OmpA, resulting in the
localization of the both Cex and CBDCex on the E. coli cell surface.
Cell-surface anchored CBDCex tightly bound to cellulose microbrils, while Cex successfully hydrolyzed the chromogenic substrate
p-nitrophenyl-b-D-cellobioside. Recently, Thermobida fusca BGL
was displayed on the E. coli cell surface using the Blc anchor
protein (Tanaka et al., 2011). The recombinant strain demonstrated
direct growth on cellobiose and cellooligosacchride. As the binding
of recombinant E. coli to cellulosic materials enables the recovery
of cells by separating the biomass from the fermentation supernatant, cellulase-immobilized cell biocatalysts have clear advantages
for CBP.
To achieve the CBP of lignocellulosic materials, genetically engineered E. coli strains were used for the production of ethanol from
sugar beet pulp (SBP) composed of pectin, cellulose, and hemicellulose. Edwards et al. (2011) successfully produced ethanol from
SBP by conferring cellobiose-fermenting and pectin-degrading
abilities to recombinant E. coli strain KO11, which stably expresses
chromosomally integrated pyruvate decarboxylase (pdc) and alcohol dehydrogenase II (adhB) genes from Zymomonas mobilis. To allow strain KO11 to utilize both cellobiose and pectin, which
contains partially methylesteried 1,4-linked a-D-galacturonic
acids, as carbon sources, the Klebsiella oxytoca cellobiose phosphotransferase operon (casAB), encoding a cellobiose permease and
phospho-b-glucosidase, Erwinia chrysanthemi pectate lyase (PelE)
gene, E. chrysanthemi out operon, which is involved in the Secdependent pathway for the secretion of PelE, and E. chrysanthemi
ogl gene, encoding an oligogalacturonide lyase, were integrated
into the KO11 genome. Following the addition of commercially
available cellulase and pectin methylesterase, the engineered
E. coli strain showed improved ethanol production from SBP due
to the effective hydrolysis of the methylester moiety of pectin by
pectin methylesterase (Edwards et al., 2011). Although additional
Table 1
Production of chemicals from cellulosic materials with non-cellulolytic bacteria in SSF and CBP processes.
Strain
B. coagulans 36D1
Relevant genes
None
Substrate
96 g L
Enzyme added
cellulose
A. aculeatus bglucosidase
None
55 g L 1 IL-treated
switchgrass
None
71 mg L
55 g L 1 IL-treated
switchgrass
None
28 mg L 1
n-butanol
55 g L 1 IL-treated
switchgrass
None
1.7 mg L
pinene
A. aculeatus bglucosidase
A. aculeatus bglucosidase
B. coagulans DSM2314
None
B. coagulans P4-102B
B. subtilis168
C. glutamicum pCCT-engD
C. cellulovorans engD
7.0 g L 1 amorphous
cellulose
15 g L 1 barley b-glucan
Carboxymethyl cellulose
E. coli MG1655
Reference
80 g L L(+)lactic acid
39.6 g L 1 L(+)lactic acid
2.0 g L 1 h 1 D(-)
lactic acid
3.1 g L 1 L(+)lactic acid
0.17 g L 1 Lglutamic acid
N.D.
2gL
cellohexaose
2gL
barley b-glucan
15 FPU g-cellulose
biocellulase W
98 mg g-biomass 1
cellulase GC220
7.5 FPU g-cellulose
cellulase
None
Product
1
FAEE
Ou et al.
(2011)
Maas et al.
(2008)
Wang et al.
(2011)
Zhang et al.
(2011b)
Tsuchidate
et al. (2011)
Hyeon et al.
(2011)
Bokinsky et al.
(2011)
Bokinsky et al.
(2011)
Bokinsky et al.
(2011)
Okano et al.
(2010)
Okano et al.
(2010)
516
3.4. C. glutamicum
C. glutamicum is a non-pathogenic, non-sporulating, non-motile,
Gram-positive soil bacterium belonging to the order Actinomycetales, which includes species of Corynebacteria, Nocardia, and Rhodococci. C. glutamicum is an important industrial microorganism due
to its high production of amino acids, which are widely used in
medicine, fodder, and as food supplements. Genetically engineered
strains of C. glutamicum are also superior to other microorganisms
for producing various kinds of organic compounds, including ethanol, 1,5-diaminopentane (cadaverine) as a component of bio-based
nylons, and succinic acid as a polymer building block (Verts et al.,
2012). Current processes for the industrial production of amino
acids and building blocks for organic polymers with C. glutamicum
predominantly focus on media supplemented with glucose, fructose, and sucrose, because wild-type C. glutamicum lacks the ability
to utilize pentose sugars, including xylose and arabinose, and cellobiose. As most C. glutamicum strains also lack the EG and BGL
activities necessary to hydrolyze plant cellulose, many attempts
have been made to construct C. glutamicum strains capable of utilizing biomass-derived sugars.
To identify suitable strains for cellobiose fermentation, the
screening for spontaneous mutant strains of C. glutamicum R capable of growing on cellobiose as the sole carbon source was performed (Kotrba et al., 2003). One mutant was identied with a
single mutation in codon 317 of bglF, which is part of the b-glucoside utilization operon consisting of the genes bglF, bglA, and bglG,
encoding a b-glucoside-specic IIBCA component of the
phosphoenolpyruvate: sugar phosphotransferase system, phospho-b-glucosidase, and anti-terminator protein from the BglG/SacY
family of transcription regulators, respectively. To allow simultaneous utilization of the sugars glucose, xylose, and cellobiose, ve
copies of clusters of xylA and xylB, which encode xylose isomerase
and xylulokinase (XK), respectively, from E. coli and one copy of a
variant bglF317A-bglA cluster from the spontaneous C. glutamicum
mutant were integrated into the genome of C. glutamicum R (Sasaki
et al., 2008). The resulting strain, X5C1, completely consumed
40 g L 1 glucose, 20 g L 1 xylose, and 10 g L 1 cellobiose within
12 h and predominantly produced lactic and succinic acids under
growth-arrested conditions. Recently, the expression of xylA and
xylB from E. coli in C. glutamicum yielded cadaverine from
517
duced 90 g L 1 L(+)-lactic acid from 100 g L 1 cellobiose at a productivity rate of 2.25 g L 1 h 1 (Adsul et al., 2007). More recently,
UV mutagenesis of Lactobacillus lactis NCIM 2368 led to the isolation of mutant strain RM2-24, which was able to produce high
amounts of D-lactic acid (110 g L 1) from 150 g L 1 acid hydrolysate of sugar cane containing cellobiose (Joshi et al., 2010).
For the efcient fermentation of lignocellulosic hydrolysates,
microorganisms must utilize not only cellobiose, but also pentoses,
particularly xylose and arabinose, present in the material. In some
LAB, arabinose and xylose are converted to xylulose-5-phosphate
(X5P) by a number of reaction steps (Okano et al., 2010). X5P is
converted to equimolar amounts of lactic acid and acetic acid via
the phosphoketolase (PK) pathway, which is an obstacle to highyield lactic acid production. In Lactococcus lactis, X5P is assimilated
by the PP pathway that produces only lactic acid as a nal product
in addition to the PK pathway. Thus, purity of D-lactic acid was improved by the redirection of the PK pathway to the PP pathway in a
recombinant Lactobacillus plantarum DldhL1 strain by substituting
an endogenous PK gene with L. lactis transketolase gene involved
in the PP pathway (Okano et al., 2010). Okano et al. (2010)
successfully produced homo-D-lactic acid from several components of lignocellulosic hydrolysate, including xylose, and
cellooligosaccharide, using metabolically engineered strains of L.
plantarum. To more effectively ferment cellulosic material, EG from
C. thermocellum has been expressed in L. plantarum (Bates et al.,
1989) and the probiotic lactobacilli Lactbacillus gasseri and
Lactbacillus johnsonii (Cho et al., 2000). Secretion of CelA by the
CelA-secreting DldhL1 strain led to the production of 1.27 g L 1
1
D(-)-lactic acid with an optical density of 99.5% from 2 g L
of
cellohexaose. Moreover, by adding BGL from A. aculeatus to the
medium of the modied strain, D(-)-lactic acid was directly produced from b-glucan (Okano et al., 2010). Narita et al. (2006) developed a cell surface display system in Lactobacillus casei using the B.
subtilis PgsA anchor protein. The display of cellulolytic enzymes on
the cell surface of LAB would enable efcient lactic acid production
from cellulosic materials.
Expression of cellulosomes in LAB strains would be a signicant
step towards the realization of the CBP of plant biomass. Recently,
a prototypic cellulosome-displaying LAB strain was constructed by
functionally binding BGL fused with the type 1 dockerin motif of
the cellulosomal enzyme, CelS, to the scaffolding protein CipA from
C. thermocellum (Wieczorek and Martin, 2010). The authors conrmed that the cellulosome-inspired enzyme complexes were displayed on the cell surface of Lactococcus lactis. BGL from Fibrobacter
succinogenes was also displayed on the cell surface of a probiotic
Lactobacillus reuteri strain (Huang et al., 2011). The construction
of mini-cellulosomes on L(+)- or D(-)-lactic acid-producing LAB
strains represents a promising platform for the production of bioplastics from cellulosic materials, and will likely be achieved in the
near future.
3.6. S. cerevisiae
The yeast S. cerevisiae is one of the most well-studied organisms
for ethanol fermentation from cellulosic and hemicellulosic materials. In addition, S. cerevisiae displays several advantageous characteristics for use in industrial applications, including its inherent
resistance to low pH, high temperature, and various inhibitors
(Hasunuma and Kondo, 2011). Accordingly, numerous studies have
described CBP using cellulase-expressing S. cerevisiae. Notably,
yeast cell-surface display technology has been widely used to
introduce cellulose-degrading abilities to S. cerevisiae. Hasunuma
and Kondo (2011) have recently reviewed lignocellulolytic ethanol
production by cell surface-engineered S. cerevisiae; therefore, here,
we have only briey described a few notable examples of CBP
using engineered S. cerevisiae.
518
519
C. cellulolyticum pWH320
Relevant genes
Z. mobilis adhII, pdc
Dldh, Dmdh
C. japonicas MSB280
C. thermocellum M1570
high-yield production of target compounds by CBP, but the construction of stable transgenic strains has been limited by the lack
of effective genetic tools. Tripathi et al. (2010) developed positive
and negative selectable markers to allow for the selection of strains
that undergo allele replacement in the absence of replicating plasmids. As C. thermocellum is sensitive to the anti-metabolite 5-uoroorotic acid, the gene encoding PyrF, which participates in de novo
pyrimidine biosynthesis, was used as a marker for selection or
counter selection. This technology was applied to delete a gene involved in acetic acid formation (pta), which improved ethanol
yields (Tripathi et al., 2010). Argyros et al. (2011) created a counter-selection system based on endogenous hpt and Thermoanaerobacterium saccharolyticum tdk genes. Combining these selection
markers enabled the use of replicating plasmids to insert and remove markers from the host chromosome. Following the deletion
of the L-lactate dehydrogenase (ldh) and pta genes from C. thermocellum with the counter-selection system and a 2000-h adaptation
period, the ethanol yield of the mutant increased 4.2-fold compared to wild-type cells. Co-culture of the C. thermocellum mutant
and T. saccharolyticum for 146 h yielded 38.1 g L 1 ethanol from
92.2 g L 1 Avicel microcrystalline cellulose and concentrations of
acetic and lactic acids that were below detection limits.
One of the challenges for the industrial application of Clostridium species is increasing its low ethanol tolerance, which typically
ranges from 1020 g L 1 in wild-type strains, as this would have a
signicant effect on lowering production costs due to the reduced
size of the necessary equipment, such as fermentation tanks and
distillation columns (Mussatto et al., 2010). Using an evolutionary
engineering approach, Shao et al. (2011) isolated ethanol-tolerant
strains of C. thermocellum that were capable of growing in medium
containing up to 50 g L 1 ethanol. Analysis of the genome sequences of the ethanol-tolerant strains revealed six common
mutations, which were found in genes involved in ethanol, arginine, and pyrimidine biosynthesis pathways. Brown et al. (2011)
revealed that a shift of cofactor specicity from NADH to NADPH
for the bi-functional acetaldehyde-CoA/alcohol dehydrogenase enzyme affords ethanol tolerance to C. thermocellum, likely by affecting electron ow.
4.4. Clostridium phytofermentans and other cellulolytic bacteria
C. phytofermentans, which has the highest number of genes
associated with the degradation of lignocellulosic material among
sequenced clostridial species (Weber et al., 2010), is one of the
promising native cellulolytic microbes for CBP. Similar to fungi,
C. phytofermentans secretes non-complexed enzymes for the
hydrolysis of lignocellulosic materials, including cellulose, hemicellulose, and pectin, to fermentable sugars. In addition, C. phytofermentans can consume nearly all of the sugars present in
Substrate
50 g L
cellulose
10 g L Avicel microcrystalline
cellulose
10 g L 1 acid-pretreated
switchgrass
10 g L 1 crystalline cellulose (Sigma
type 50)
10 g L 1 Avicel microcrystalline
cellulose
20 g L 1 Avicel microcrystalline
cellulose
Product
Reference
1
0.83 g L
ethanol
2.7 g L 1
ethanol
1.3 g L 1
ethanol
0.42 g L 1
isobutanol
0.0035 g L
ethanol
5.6 g L 1
ethanol
lignocellulosic biomass, such as arabinose, cellobiose, fructose, galactose, gentiobiose, glucose, lactose, maltose, mannose, ribose, and
xylose, to produce ethanol and acetate as the major products
(Weber et al., 2010). In 10-day fermentation experiments with
AFEX-treated corn stover containing 0.5% (w/w) glucan,
C. phytofermentans hydrolyzed 76.0% and 88.6% of available glucan
and xylan, respectively, and produced 2.8 g L 1 ethanol. The ethanol yield achieved by the CBP with C. phytofermentans was 71.8%
of those obtained by SSCF using commercially available cellulase
and a xylose-fermenting S. cerevisiae strain (Jin et al., 2011).
Numerous microorganisms, including hyperthermophilic marine archaea, which can tolerate elevated temperatures are able to
utilize carbohydrates such as a- and b-linked glucan, xylan, and
mannan for growth, whereas the utilization of crystalline cellulose
is limited to a number of terrestrial bacteria with upper temperature limits for growth of below 75 C (Blumer-Schuette et al.,
2008). Anaerocellum thermophilum is a Gram-positive thermophilic
anaerobe that grows optimally at 75 C. A. thermophilum strain
DSM 6725 (Caldicellulosiruptor bescii) efciently utilizes various
types of untreated plant biomass, including hardwoods, grasses,
crystalline cellulose, and xylan, with the predominant end-products being hydrogen, acetate, and lactate (Lochner et al., 2011).
Caldicellulosiruptor saccharolyticus, which is a close relative of A.
thermophilum, can degrade crystalline cellulose (Blumer-Schuette
et al., 2008). Interestingly, C. saccharolyticus strain DSM 8903 can
grow on switchgrass, but not on poplar. Yang et al. (2009) demonstrated a signicant difference in the biomass hydrolysis activity
between A. thermophilum DSM6725 and C. saccharolyticus
DSM8903, which would be due to the recalcitrance of poplar for
the DSM8903 strain.
C. thermocellum exhibits a high growth rate on crystalline cellulose, but does not utilize xylan and cannot grow on xylose or other
pentoses. However, hemicellulolytic thermophiles could be used in
conjunction with C. thermocellum to hydrolyze hemicellulose and
ferment all of the sugars present in the biomass substrate (Shaw
et al., 2008). The thermophilic anaerobe T. saccharolyticum is able
to utilize xylan and biomass-derived sugars, including arabinose,
mannose, and xylose, as well as glucose, as carbon sources to produce ethanol and organic acids. In order to improve ethanol yields,
Shaw et al. (2008) knocked out the genes involved in organic acid
formation, such as acetate kinase, phosphate transacetylase, and Llactate dehydrogenase, which resulted in a T. saccharolyticum
strain capable of producing ethanol as the only detectable product.
Using the homoethanologenic strain for the SSF of Avicel at 50 C
resulted in a 2.5-fold reduction in cellulase loading compared to
that required for S. cerevisiae cultured at 37 C (Shaw et al.,
2008). Recently, cellulose- and xylan-degrading thermophilic
anaerobes isolated from self-heated biocompost were able to utilize xylan with roughly the same efciency as that for cellulose
520
(Sizova et al., 2011). Phylogenetic analysis of 16S rRNA and glycosyl hydrolase family 48 gene sequences suggested that the isolates
were related to a strain of Clostridium clarifravum.
Establishing economically feasible fermentation processes requires markedly increasing nal product titers due to the high energy demands of subsequent product recovery steps, as well as the
capital and production costs associated with biorenery equipment (Hasunuma and Kondo, 2011; Mussatto et al., 2010).
Although high-yield production of target compounds by metabolically optimized microbes is necessary, achieving higher titers inevitably requires increased loading of solid lignocellulose in the SSF
and CBP processes. Increase in the solid concentration results in
corresponding increases in chemical production (Matano et al.,
2012). However, by increasing the lignocellulose content in the
bioreactor, the concentration of fermentation inhibitors released
during the pretreatment of biomass would reach higher levels.
Accordingly, microorganisms that are resistant to inhibitors are a
prerequisite for the high-titer production of fuels and chemical
products (Hasunuma and Kondo, 2011).
To further engineer cellulolytic recombinant and native strains
for use in CBP, system-wide modications of intracellular metabolic pathways using advanced engineering tools such as minimal
hosts, vectors, genetic controllers, and characterized enzymes
(Keasling, 2012) are needed, which would improve the potential
of not only target productivities but cell growth and viability during the fermentation. In the near future, it may be possible to design optimized metabolic systems with the aid of computer
simulations based on metabolic proling data (Kondo et al.,
2012a). The integration of cellulolytic capabilities with metabolic
systems specied for targeted chemical production will allow customized CBP microorganisms to be developed using advanced gene
manipulation technologies.
7. Conclusions
5. Synthesis
Recent progress in the development of microorganisms through
evolutionary, metabolic, and synthetic engineering approaches
have paved the way for utilizing lignocellulosic biomass as a bioresource for fuel and chemical production. However, the one-step
fermentative conversion of cellulosic material to chemical products represents a major challenge to achieving this goal. When
using recombinant cellulose-utilizing microbes, low cellulolytic
activities of heterologous enzymes often result in unutilized cellulose, thereby decreasing yields. One major obstacle to modifying
non-cellulolytic microorganisms for the utilization of lignocellulose is the low inherent capacity of these strains for foreign cellulase secretion (Bokinsky et al., 2011). To overcome this limitation,
protein export systems that do not compromise the cell membrane
should be introduced into recombinant strains. In addition, as the
efcient degradation of crystalline cellulose requires the synergistic reaction of cellulases, appropriate combinations and quantitative ratios of cellulase subsets need to be expressed in strains
used for the fermentation of lignocellulosic biomass (Yamada
et al., 2011). As an alternative approach, cellulase engineering,
which has offered improvement in specic activity at a broad range
of pH and temperature by directed evolution and rational design
(Chandel et al., 2012), might be implemented. When using native
cellulose-utilizing microbes for industrial applications, these
microorganisms should exhibit high product yields, productivities,
and product titers, while not over-burdening cells energetically.
Although genetic manipulation tools for the microbes are under intense development, the ability to engineer complete metabolic
In this review, recent advances in the development of microorganisms for the fermentative production of bio-based fuels and
chemicals from cellulosic materials were highlighted. Direct conversion of cellulose to key commodities such as alcohols, fatty
acids, isoprenoid and organic acids has been achieved through
the engineering of metabolic pathways and cellulose hydrolysis
abilities of microorganisms. Despite of the obvious requirements
for increasing yields and lowering production costs, this signicant
proof-of-concept will allow the application of the engineered
microorganisms to advancing CBP technologies.
Acknowledgements
This work was supported by project P07015 of the New Energy
and Industrial Technology Development Organization (NEDO) under the sponsorship of the Ministry of Economy, Trade, and Industry (METI) of Japan. This work was also supported by Grants-in-Aid
for Young Scientists (B) to TH and KYH from the Ministry of Education, Culture, Sports and Technology (MEXT) of Japan and Special
Coordination Funds for Promoting Science and Technology, Creation of Innovative Centers for Advanced Interdisciplinary Research
Areas (Innovative Bioproduction Kobe), MEXT, Japan.
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