Review

Biotechnological production
of succinic acid: current state
and perspectives
Ke-Ke Cheng, Xue-Bing Zhao, Jing Zeng and Jian-An Zhang, Tsinghua University, Beijing, P.R. China

Received November 23, 2011; revised December 15, 2011; accepted December 16, 2011
View online February 29, 2012 at Wiley Online Library (wileyonlinelibrary.com); DOI: 10.1002/bbb.1327;
Biofuels, Bioprod. Bioref. 6:302–318 (2012)

Abstract: Succinic acid has multiple practical applications (e.g. synthesis of 1,4-butanediol, tetrahydrofuran,
gamma-butyrolactone, and as a monomer of some biodegradable polymers). Bio-based succinic acid is a potential
substitute for current petrochemical production. Facing a shortage of crude oil supply and sharply rising oil prices,
biological production of succinic acid from abundant and available biomass has become a topic of worldwide
interest. Although great progress has been made in recent decades, much needs to be developed further in order to
achieve economic viability. This paper reviews developments in technology and updates research progress of bio-
succinate production, including pathways, micro-organisms, culture conditions, as well as integrated production with
other high-value-added products. Finally, strategies are proposed for successful commercialization of fermentative
succinic acid production. © 2012 Society of Chemical Industry and John Wiley & Sons, Ltd

Keywords: bioconversion; biomass; fermentation; succinic acid

Introduction and plant growth. Based upon the structure of linear and
saturated dicarboxylic acid, succinic acid can be readily
uccinic acid (IUPAC name: butanedioic acid) is converted to other bulk chemicals, such as 1, 4-butanediol,

S a colorless crystal soluble in water, ethanol, and

acetone. It is also known as amber acid since it was
first obtained by distilling amber in 1550. Succinic acid has
gamma-butyrolactone, tetrahydrofuran, adipic acid,
n-methylpyrrolidone or linear aliphatic esters.1-4 With vari-
ous environmental implications, the demand for succinic
a wide range of industrial applications, such as being used acid is expected to increase significantly. For example, a
as a chemical intermediate for the production of lacquers new biodegradable polymer, poly(1,3-propylene succinate),
and perfume esters as well as a flavor, bacteriostatic, or can be derived by thermal polycondensation of succinic
neutralizing agent in the food industry. Furthermore, suc- acid with 1,3-propanediol.5 Succinic acid can be converted
cinic acid also has a special chemical market for produc- to 1,4-butanediol,6-8 which can be further used for the
ing coatings, surfactants, dyes, detergents, green solvents, production of biodegradable poly(butylene succinate) (PBS)
biodegradable plastics, and ingredients stimulating animal with excellent thermal and mechanical properties as well

Correspondence to: Jian-An Zhang, Institute of Nuclear and New Energy Technology, Tsinghua University, Beijing 100084, P.R. China.

E-mail: zhangja@tsinghua.edu.cn

302 © 2012 Society of Chemical Industry and John Wiley & Sons, Ltd
Review: Biotechnological production of succinic acid K-K Cheng et al.

as thermoplastic processability.9 While the current global Mitsubishi Chemicals, and PTT have their individual plans
succinic acid production is 30 000 to 50 000 tons per year for commercial bio-based production of succinic acid.8
with the market price of US$2400–3000 per ton, the market Recognizing the importance of the biotechnological pro-
is expected to reach 100 000 tons per year by 2015.1,3,10 duction of succinic acid, herein we intend to review relevant
Succinic acid can be produced via chemical routes, which synthetic pathways, micro-organism producers, cultivation
mainly include paraffi n oxidation,11 catalytic hydrogena- technologies, as well as propose effective strategies toward
tion, or electroreduction of maleic acid or maleic anhy- bioprocess commercialization through the integrated produc-
dride.12,13 In the paraffi n oxidation process, a calcium tion of succinic acid with other high-value-added products.
or manganese catalyst is employed and several kinds of
dicarboxylic acids are obtained at the same time. Succinic Metabolic pathway
acid is then recovered and purified by distillation, crys- Being a key intermediate of the tricarboxylic acid (TCA)
tallization, and drying. However, the yield and purity of cycle, succinic acid can also act as a fermentation end-
succinic acid obtained by this process are relatively poor.11 product for a selection of bacterial strains when glucose or
Industrialized as early as the 1930s, the hydrogenation glycerol is used as a carbon source with the fi xation of car-
process is one of the mature technologies for chemical bon dioxide. There are three routes that can form succinate:
production of succinic acid. The reaction can be conducted the reductive branch of the TCA cycle, also known as the
homogeneously or heterogeneously by careful selection fermentative pathway (which is primarily active under fully
of the catalyst. While high yield, purity, and selectivity of anaerobic conditions), the oxidative branch of the TCA cycle
succinic acid can be obtained, the operation of this process (which is primarily active under aerobic conditions), and the
is complicated, expensive, and might have environmental glyoxylate pathway, which is essentially active under aerobic
implications.13 conditions upon adaptation to growth on acetate.
Recent developments in the production of succinic acid Depending on the stoichiometric assimilation of car-
have been focused on biotechnological alternatives, in bon dioxide and hydrogen, the overall yield can vary
particular microbial transformation based upon the use of significantly: 4,10,18
renewable biomass as feedstock.14-17 Note that the fact that
CO2 is assimilated during succinic acid fermentation can be C6H12O6 + CO2→ HOOCCH2CH2COOH + CH3COOH 
+ HCOOH
considered as an environmental advantage.
7C6H12O6 + 6CO2 → 12 HOOCCH2CH2COOH + 6 H2O
At present, the biotechnological production of succinic
C6H12O6 + 2CO2 + 2H2 → 2HOOCCH2CH2COOH + 2H2O
acid is still at demonstration scale, but significant progress
is expected in light of major business joint ventures and Under anaerobic conditions, succinate derived from phos-
research and development activities. For example, BioAmber, phoenolpyruvate (PEP), via several intermediate compounds
a US company dedicated to the production of bio-based of the TCA cycle, including oxaloacetate (OAA), malate, and
succinic acid, has constructed a demonstration plant in fumarate, is recognized as the primary pathway. Depending
Pomacle, France, with a capacity of 2000 tons per year. In on the micro-organism and cultivation condition, other
parallel, BioAmber developed a turn-key technological metabolites, such as ethanol, acetate, lactate, and formate, can
package available for licensing in 2011. DSM and Roquette be synthesized when pyruvate is further oxidized.19-20 The
are currently building a large-scale plant with a capacity of fermentative pathway converts oxaloacetate to malate, fuma-
10 000 tons per year and will begin commercial production rate, and then succinate and this pathway requires 2 moles of
in 2012. Myriant, a successor to BioEnergy International, NADH per mole of succinate produced. One major obstacle
was recently awarded US$50 million by the US Department to high succinate yield through the fermentative pathway
of Energy to construct a succinic acid plant with an initial is due to NADH limitation. This is because 1 mole glucose
capacity of 13 500 tons per year in Louisiana, USA, which is can provide only 2 moles of NADH through the glycolytic
expected to be running in 2012. In addition, BASF, Purac, pathway; however, the formation of 1 mole succinate through

© 2012 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 6:302–318 (2012); DOI: 10.1002/bbb 303
K-K Cheng et al. Review: Biotechnological production of succinic acid

the native fermentative pathway requires 2 moles NADH. During anaerobic culture, E. coli undergoes mixed-acid
Therefore, the maximum molar theoretical yield of succi- fermentation that yields acetate, ethanol, formate, and lactate
nate from glucose is limited to 1 mol mol-1 glucose assuming as its major fermentation products with only a small amount
that all the carbon flux will go through the native succinate of succinate formed. The primary pathway for succinate
fermentative pathway. In the bacterium Actinobacillus suc- synthesis occurs by carboxylation of PEP to oxaloacetate
cinogenes, glucose is metabolized to PEP by glycolysis and using phosphoenolpyruvate carboxylase, which is irreversible
the oxidative pentose phosphate pathway.20,21 To further (Fig. 2). Different from A. succinogenes, conversion of PEP to
synthesize succinate from PEP, four key enzymes, including oxaloacetate using phosphoenolpyruvate carboxykinase is
phosphoenolpyruvate carboxykinase, malate dehydrogenase, reversible and usually active during gluconeogenesis. Another
fumarase and fumarate reductase are required (Fig. 1). potential biosynthetic route for succinate in E. coli is through
In E. coli, succinate is a metabolite that is formed under the glyoxylate pathway, which is active under aerobic condi-
both anaerobic and aerobic conditions. Under aerobic condi- tions. The glyoxylate pathway converts 2 mol acetyl CoA and
tions, acetyl-CoA generated from pyruvate mainly enters the 1mol OAA to 1mol succinate and 1mol malate, which can be
TCA for energy and cell intermediates production. Succinate further converted to succinate using only 1mol NADH. The
is formed by succinyl-CoA synthetase and subsequently con- IclR transcriptional repressor (encoded by the gene iclR), reg-
verted to fumarate by succinate dehydrogenase. As a result, ulates the expression of the aceBAK operon involved in the
succinate does not accumulate in wild-type E. coli cultures induction of the glyoxylate pathway upon growth on acetate
under aerobic condition. To realize succinate aerobically under aerobic conditions. By deleting the iclR, the glyoxylate
accumulation, inactivation of sdhA gene to block the con- pathway will be activated in anaerobic condition, which will
version of succinate to fumarate in TCA cycle is necessary.22 benefit to achieve higher succinate yield.
The yield of succinate from sugars or other carbon sources
is strongly decided by available NADH and ATP produced in
the C3 route which results in by-product accumulation.23,24
In view of the high cost of product recovery, a high final prod-
uct concentration with high yield is desirable. To achieve this,
the formation of acetate, lactate, formate, and ethanol should
be controlled at the minimum levels.25,26 The metabolic
pathway indicates that by-product formation is the result of
carbon distribution among phosphoenolpyruvate–pyruvate
node. The flux toward pyruvate should be controlled so that
it allows necessary pyruvate as a biosynthetic precursor.
Metabolic engineering can be very helpful in manipulating
the pathways so that by-product pathways will be controlled
or even eliminated. Changing the activity of enzymes around
PEP, pyruvate and OAA would also entail flux redistribution,
Figure 1. Succinic acid production pathway in A. succinogenes
which could bring improvement in the succinate synthesis.
(simplified, based on van der Werf et al.20). (1) Embden–Meyerhof
pathway enzymes; (2) pentose phosphate pathway enzymes; (3)
phosphoenolpyruvate carboxykinase; (4) malate dehydrogenase;
Micro-organisms
(5) fumarase; (6) fumarate reductase; (7) pyruvate kinase; (8) lactate Natural succinate producing strain
dehydrogenase; (9) oxaloacetate decarboxylase; (10) malic enzyme; Many natural bacterial species have the pathway for convert
(11) pyruvate–formate lyase; (12) acetate kinase; (13) phospho- PEP to succinate. Experimental data infer the presence of the
transacetylase; (14) acetaldehyde dehydrogenase; (15) alcohol succinic acid pathway (in whole or in part) in the following
dehydrogenase. species: Actinobacillus succinogenes,27-28 Anaerobiospirillum

304 © 2012 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 6:302–318 (2012); DOI: 10.1002/bbb
Review: Biotechnological production of succinic acid K-K Cheng et al.

Glucose
NAD+
1
NADH

Phosphoenolpyruvate
2 NADH NAD
+

3
NAD(P)H Pyruvate Lactate
CO2 CO
+ 2 4

2
9

CO
NAD(P)
10 18 Formate H2+CO2
17
5 6
Acetyl-CoA Acetyl-phosphate Acetate
NADH NADH
NAD+ Oxaloacetate 7 NAD+
+
11 16 NADH NAD
8
Acetylaldehyde Ethanol
Malate Citrate
20 Acetyl-CoA
12 15
Glyoxylate 19
Fumarate Isocitrate

NADH
13 NAD+ 14
Succinate

Figure 2. Succinic acid production pathway in E. coli (simplified, based on Jantama

et al.).17 (1) Embden–Meyerhof pathway enzymes; (2) pyruvate kinase; (3) lactate
dehydrogenase; (4) pyruvate–formate lyase; (5) phospho-transacetylase; (6) acetate
kinase; (7) acetaldehyde dehydrogenase; (8) alcohol dehydrogenase; (9) malic enzyme;
(10) phosphoenolpyruvate carboxylase; (11) malate dehydrogenase; (12) fumarase;
(13) fumarate reductase; (14) isocitrate lyase; (15) aconitase; (16) citrate synthase; (17)
oxaloacetate decarboxylase; (18) PEP carboxylase; (19) isocitrate lyase; (20) malate
synthase.

succiniciproducens,29 Bacteroides amylophilus,30 Bacteroides Genetically modified strain

fragilis,31-33 Bacteroides succinogenes,34,35 Clostridium thermo- Although A. succinogenes, A. succiniciproducens, M. suc-
succinogenes,36 Cytophaga succinicans,37 Escherichia coli.,18,38 ciniciproducens are well known succinate producers, not
Fibrobacter succinogenes,39 Klebsiella pneumoniae,40 Klebsiella many reports have been published on metabolic engineer-
oxytoca,41 Mannheimia succiniciproducens,42 Paecilomyces ing of these species.61,66 Due to a rich set of genetic tools
varioti,43 Penicillium simplicissimum,44 Ruminococcus flave- available and the fact of fast cell growth and simple culture
faciens,45 and Succinivibrio dextrinosolvens.46 It is worth not- medium, E. coli is one of the most wholly studied systems
ing that not all species produce succinate as an end-product for succinate production. Strategies in metabolic engi-
and sometimes succinate as an intermediate can be further neering of E. coli can be classified as four main methods:
converted to another product, such as propionate.31 Some improvement of substrate transportation, enhancement
publications report yeast can synthesize succinate. However, of pathways directly involved in the succinate production,
the productivity is very poor. 47-49 deletion of pathways involved in by-product accumulation,
Currently, high efficient producers of succinic acid are and their combinations.67-76 These four methods have been
Actinobacillus succinogenes,50-55 Anaerobiospirillum succinic- studied in many reports and some high efficient succinic
iproducens,56-59 and Mannheimia succiniciproducens.60-62 acid producers have been constructed.77-84 Succinic acid
Corynebacterium glutamicum has been recognized recently production using different bacteria species is compared in
to be a very important organism for succinate prodction.63-65 Table 1.

© 2012 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 6:302–318 (2012); DOI: 10.1002/bbb 305
K-K Cheng et al. Review: Biotechnological production of succinic acid

Table 1. Microbial succinic acid production using different bacteria species.

Strains Microbial succinate production References
Substrates Methods Concentration Productivity Yield
(g l-1) (g l-1 h-1) (g g-1)
A. succinogenes
FZ53 Glucose Anaerobic, batch 105.8 1.36 0.83 27
FZ6 Corn fiber hydrolyzate Anaerobic, batch 70.6 0.7 0.88 27
130Z Sake lees Anaerobic, batch 48 0.94 0.75 85
130Z Whey Anaerobic, batch 21.5 0.44 0.57 86
130Z Glucose Anaerobic, batch 67.2 0.8 0.7 87
130Z Glucose Anaerobic, batch 45.8 1.55 0.83 87
CGMCC 2650 Corn straw Anaerobic, batch 15.8 0.62 1.23 55
CGMCC 1593 Cane molasses Anaerobic, batch 50.6 0.84 0.8 88
CGMCC1593 Corn stover hydrolyzate Anaerobic, fed-batch 53.2 1.21 0.83 89
CGMCC 1593 Glucose Anaerobic, batch 60.2 1.3 0.75 90
CGMCC 1716 Corn fiber hydrolyzate Anaerobic, batch 35.4 0.98 0.73 91
CIP 106512 Sugarcane bagasse Anaerobic, batch 22.5 1.01 0.43 92
hydrolyzate
A. succiniciproducens
ATCC53488 Glucose Anaerobic, continuous 83 10.4 0.88 51
culture with membrane
for cell recycling
ATCC53488 Glucose Anaerobic, batch 34.4 1.8 0.86 57
ATCC53488 Glucose Anaerobic, batch 32.2 1.2 0.99 59
ATCC53488 Glucose Anaerobic, batch 50.3 2.1 0.9 93
ATCC53488 Glucose Anaerobic, continuous 39.1 2.03 0.85 94
culture
ATCC29305 Whey Anaerobic, fed-batch 34.7 1.02 0.91 95
ATCC29305 Whey Anaerobic, continuous 19.8 3 0.64 95
culture
ATCC29305 Glucose Anaerobic, continuous 14.3 3.3 0.71 58
culture with membrane
for cell recycling
ATCC29305 Glycerol Anaerobic, batch 19 0.15 1.6 96
ATCC29305 Glucose/Glycerol Anaerobic, batch 29.6 1.35 0.97 96
ATCC29305 Wood hydrolyzate Anaerobic, batch 24 0.74 0.88 97
ATCC29305 Galactose Anaerobic, batch 15.3 1.46 0.87 97
M. succiniciproducens
MBEL55E Glucose Anaerobic, batch 14 1.87 0.70 42
MBEL55E Whey Anaerobic, batch 13.4 1.18 0.71 60
MBEL55E Whey Anaerobic, continuous 6.4 3.9 0.69 60
culture
MBEL55E Glucose Anaerobic, batch 10.5 1.75 0.59 99
MBEL55E Wood hydrolyzate Anaerobic, batch 11.7 1.17 0.56 100
MBEL55E Wood hydrolyzate Anaerobic, continuous 8.2 3.19 0.55 100
culture
LPK7 Glucose Anaerobic, fed-batch 52.4 1.75 0.76 61
E. coli
NZN111 Glucose Anaerobic, batch 12.8 0.29 0.64 101
NZN111 Glucose Dual phase aeration, 28 0.7 0.7 102
Fed-batch
AFP111 Glucose Dual phase aeration, 51 0.52 0.54 67
batch

306 © 2012 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 6:302–318 (2012); DOI: 10.1002/bbb
Review: Biotechnological production of succinic acid K-K Cheng et al.

Table 1. Continued.
Strains Microbial succinate production References
Substrates Methods Concentration Productivity Yield
(g l-1) (g l-1 h-1) (g g-1)
AFP111 Glucose Dual phase aeration, 101 1.18 0.78 103
batch
AFP111-pyc Glucose Dual phase aeration, 99.2 1.31 1.1 104
batch
AFP184 Glucose Dual phase aeration, 38 1.27 0.8 105
batch
AFP184 Fructose Dual phase aeration, 30 1.01 0.7 105
batch
AFP184 Xylose Dual phase aeration, 23 0.78 0.5 105
batch
AFP184 Glucose Dual phase aeration, 77 0.71 0.75 76
fed-batch
AFP184 Softwood dilute acid Dual phase aeration, 42.2 0.78 0.72 103
hydrolyzate batch
KJ060 Glucose Anaerobic, batch 86.5 0.9 0.93 18
KJ060 Glucose Anaerobic, batch 73.4 0.61 1.06 18
KJ073 Glucose Anaerobic, batch 78.8 0.82 0.79 18
KJ122 Glucose Anaerobic, batch 83 0.88 0.9 107
SBS550MG Glucose Anaerobic, Fed-batch 40 0.42 1.06 108
SBS550MGpHL314 Glucose Anaerobic, Fed-batch 40 0.42 1.1 17
HL51276k- pepc Glucose Aerobic, batch 8.3 0.16 0.72 109
HL27659k-pepc Glucose Aerobic, Fed-batch 58.3 0.72 0.62 109
JCL1208 Glucose Anaerobic, batch 10.7 0.59 0.29 110
W3110 Sucrose Dual phase aeration, 24 0.81 1.2 70
batch
W3110 Cane molasses Dual phase aeration, 26 0.87 0.52 111
batch
SD121 Corn stalk enzymatic Dual phase aeration, 57.8 0.8 0.87 112
hydrolyzate batch
C. glutamicum
R Glucose Micro-aerobic, fed-batch 23 3.83 0.19 64
with membrane for cell
recycling
R ΔldhA-pCRA717 Glucose Micro-aerobic, fed-batch 146 3.17 0.92 65
with membrane for cell
recycling
B. fragilis
MTCC1045 Glucose Anaerobic, batch 12.5 0.42 0.62 32
MTCC1045 Glucose Anaerobic, batch 20 0.83 0.57 33
P. ruminocola
ATCC 19188 Glucose Anaerobic, batch 18.9 0.52 – 87
F. succinogenes
S85 Wheat straw Anaerobic, batch 1.55 0.022 0.05 113
S85 Orange peel Anaerobic, batch 1.75 0.025 0.044 113
S. cerevisiae
AH22ura3 Glucose Aerobic, batch 3.62 0.022 0.072 49
Δ
sdh2Δsdh1Δidh1Δidp1

Currently, major research results are obtained using prepared concentration and yield are obtained in fed-batch or continu-
medium and pure sugar as feedstock. Succinate productions ous culture systems. The highest productivity, 10.4 g l-1 h-1 was
are mostly carried out in batch fermentations, while higher obtained in a continuous culture of A. succiniciproducens with

© 2012 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 6:302–318 (2012); DOI: 10.1002/bbb 307
K-K Cheng et al. Review: Biotechnological production of succinic acid

integrated membrane for cell recycling at a dilution rate of whey, 21.5 g l-1 succinate with a yield of 0.57 g g-1 whey and
0.98 h−1. Succinate concentration up to 146 g l-1 was obtained productivity of 0.44 g l-1 h-1 were obtained.86 Corn stover as
in a cell recycling fed-batch culture of C. glutamicum. In an a substrate was used for succinate fermentation by A. suc-
E. coli dual phase (aerobic/anaerobic) batch fermentation using cinogenes CGMCC1593.89 The raw material was pre-treated
sucrose as substrate the highest described yield of 1.2 g g-1 by diluted alkaline before simultaneous saccharification and
sucrose was obtained. Using lactose as inducer, E. coli SD111 fermentation so that part of the lignin could be removed.
-1
also produces succinate with a high yield of 1.1 g g glucose in Cellulose and hemicellolose were further hydrolyzed by
114
aerobic/anaerobic combined fed-batch fermentations. cellulase and cellobiase, and used as a medium in succi-
nate production. Corn stalk after steam explosion and then
Culture conditions NaOH-H2O2 pre-treatment were hydrolyzed by enzyme,
obtaining a glucose, xylose, cellubiose medium for suc-
In addition to essential nutrient components for sustaining
cinate production. Using this substrate, A. succinogenes
bacterial cell growth and physiological maintenance, other
CGMCC2650 can produce 15.8 g l-1 succinate. The authors
substrates, including major carbonaceous feedstock, carbon
dioxide, and hydrogen, as well as culture pH are also critical observed that glucose and xylose were consumed simultane-
for economic production of succinate. Using a selection of ously, while cellubiose was not used until glucose and xylose
bacterial strains, these technical issues and general cultiva- were exhausted.55
tion strategies are reviewed. A. succiniciproducens can produce succinate from whey,95
glycerol,96 pre-treated wood hydrolyzate,97 and galatose.98
In the latter study, the authors examined succinate produc-
Substrate
tion using galactose, galactose/glucose, or galactose/lactose
Feedstock represents the major cost for most bioprocesses
medium, respectively. Anaerobiospirillum succiniciproducens
for the production of bulk chemicals. Therefore, identify-
ATCC 29305 can consume galactose as the sole substrate
ing inexpensive feedstock for the production of succinate
at a rate of 2.4 g g dry cell weight-1 h-1 and synthesis suc-
becomes critical to making the bioprocess economical.
Among various feedstock options, waste by-products (such cinate in a yield of 0.87 g g-1. When glucose and galactose
as glycerol from biodiesel production) and waste biomass coexisted, A. succiniciproducens consumed two kinds of
(such as corn stover and corncob) appear to be promising sugars synchronously. Further, when a lactose and galactose
based upon many successful attempts. mixture was used as substrate, galactose consumption was
A selection of substrates has been used for growing not affected by lactose. The authors concluded that the pro-
Actinobacillus succinogenes to produce succinate, include ductivity and economy of biological succinate production
sake lees,85 cane molasses,88 corn stover,89,115,116 corn could be improved by co-fermentation from galactose and
fiber,91,117 wheat flour,118 corncob hydrolysates,119 corn stalk other sugars. Succinate production from whey, whose main
and cotton stalk,55 sugarcane bagasse hemicellulose hydro- compositions are lactose, protein, and lactic acid, was inves-
lyzate,92 and whey.86 Cane molasses, pre-treated by sulfu- tigated in batch, continuous, and fed-batch cultures using
ric acid, were consumed as carbon resource for succinate A. succiniciproducens ATCC 29305. Using a CO2 limited
synthesis using A. succinogenes CGMCC1593. In anaerobic medium (1g l-1 MgCO3), only 48% of lactose was metabo-
bottles fermentation, 50.6 g l-1 succinate with a productivity lized and lactic acid accumulated as a major product. Using
of 0.84 g l-1 h-1 was obtained, and the sugar conversion ratio a high CO2 level medium (35 g l-1 MgCO3), more than 90%
was about 95.6%. During the fed-batch fermentation with of the lactose was metabolized and succinate accumulated
external CO2 aeration and controlled pH, higher succinate as major product. The highest succinate productivity of 3 g
concentration (55.2 g l-1) and productivity (1.15 g l-1 h-1) were l-1 h-1 reached in continuous culture and the yield was 0.64
reached.88 Succinate was synthesized by A. succinogenes g g-1. Higher product concentration and yield (34.7 g l-1 and
130 Z from cheese whey. At pH 6.8, inoculum size of 5%, 0.906 g g-1) were obtained in fed-batch cultures.95 Lee et al.96
0.5 vvm CO2 aeration and 200 rpm, initial 50 g l-1 cheese found that A. succiniciproducens ATCC 29305 can metabo-

308 © 2012 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 6:302–318 (2012); DOI: 10.1002/bbb
Review: Biotechnological production of succinic acid K-K Cheng et al.

lize glycerol as a carbon source to accumulate succinate and both the pHL413 plasmid, which contains L. lactis pycA
glycerol consumption depending on the yeast extract con- gene, and the pUR400 plasmid, which contains the scrK, Y, A,
centration in the medium. When glycerol was used as the B, and R genes for sucrose uptake and catalyzation was tested
only carbon source in a medium added with yeast extract, using fructose, sucrose, a mixture of glucose and fructose, a
the highest 19 g l-1 succinate was obtained. When a glycerol- mixture of glucose, fructose and sucrose, and sucrose hydrol-
glucose mixture was used as co-substrate, 29.6 g l-1 succinate ysis solution medium for succinate production. Compared to
was produced. The authors concluded that succinate produc- the culture grown on fructose or sucrose alone, co-utilization
tion from glycerol has some advantages over glucose, such as of glucose with fructose and sucrose increased in succinate
increased succinate yield and decreased acetate formation, productivity. When SBS550MG pHL413 pUR400 was
which benefit downstream processes because acetate is not cultured using a glucose–fructose–sucrose mixture medium,
easy to separate from succinate. glucose was utilized preferentially to sucrose and sucrose was
M. succiniciproducens produced succinate when grown utilized preferentially to fructose.120
on whey60 and wood hydrolyzate.101 In the latter study, oak Sugars from waste biomass hydrolyzates (especially those
wood cellulosic residues after steam explosion pre-treatment derived from the hemicellulose fraction) have been used for
were hydrolyzed by cellulase, obtaining a glucose/xylose succinate fermentation. However, the cellulosic feedstock
mixture. This mixture was treated with sodium hydroxide often contains inhibitors which negatively affect the fermen-
before sterilization for reducing inhibitors. M. succinicipro- tation efficiency and succinate yield. These inhibitors might
ducens MBEL55E metabolized xylose/glucose co-substrate include weak acids, furans, and phenolic compounds, and
can be partly removed by detoxification process, such as
in the wood hydrolyzate-based medium for succinate pro-
treatment with overliming or activated carbon adsorption.
duction. In batch fermentations, 11.7 g l-1 succinate was
However, the fermentation parameters for the treated hydro-
obtained, giving a succinate yield of 0.56 g g-1 sugar and a
lyzate are still lower than those obtained with a synthetic
productivity of 1.17 g l-1 h-1; while the continuous fermenta-
medium. This shows that there were some leftover toxic com-
tions at a dilution rate of 0.4 h−1 obtained similar succinate
ponents in the treated hydrolyzate that negatively affected the
yield (0.55 g g-1) but higher productivity of 3.19 g l-1 h-1.
fermentation performance. To further improve the fermenta-
Using the same strain, another substrate, whey and corn
tion efficiency, proper culture conditions and detoxification
steep liquor, were investigated for succinate production in
techniques should be developed to alleviate the inhibitions.
batch and continuous fermentation. In batch fermentations,
13.4 g l-1 succinate was obtained, giving a succinate yield of CO2 supply
0.71 g g-1 lactose and a productivity of 1.18 g l-1 h-1. In the CO2 supply is an important variable in succinate fermenta-
continuous fermentations succinate yield is in the range of tion. Not only can external CO2 gas supply but also some
0.63–0.69 g g-1 lactose. The highest productivity of 3.9 g l-1 carbonates in the medium can be considered as a source of
h-1 was obtained at a dilution rate of 0.6 h−1.60 CO2. As a consequence, for a high succinate yield, CO2 or
Succinate production using corn stalk enzymatic hydro- some carbonates are needed. When CO2 or carbonates dis-
lyzate by recombinant strain E. coli SD121 was studied by solve in water, dissolved CO2 or carbonates react with water
Wang et al.112 In this case, a two-stage aeration strategy was producing HCO3- and CO3-. The equilibrium among CO2,
used. The aerobic culture was carried out for the first 12 h HCO3- and CO3- are decided by pH in medium.
with a reducing sugar concentration about 44 g l−1 and the The effects of carbon dioxide levels on the glucose fermen-
cell growth entered into the middle exponential phase with a tation and cells grown at pH 6.2 were studied by Samuelov
dry cell weight of about 7.6 g l−1. Then the anaerobic culture et al.121 In this study, CO2 source was provided by MgCO3.
was triggered by CO2 aeration. The final concentration of When the molar ratio between CO2-HCO3- and glucose
succinate was 57.81 g l−1. The overall productivity and yield of was 0.5–1.0, about 15% of the available carbon (glucose plus
succinate in the whole anaerobic stage were 0.96 g l −1 h−1 and CO2-HCO3-) was converted to biomass. Sixty-five percent
0.87 g g−1 total sugar, respectively. E. coli SBS550MG bearing of the carbon was converted to succinate. When the molar

© 2012 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 6:302–318 (2012); DOI: 10.1002/bbb 309
K-K Cheng et al. Review: Biotechnological production of succinic acid

ratio between CO2-HCO3- and glucose was 0.065, only 8% of MBEL55E.123 Biomass formation was strongly inhibited
the carbon was converted into biomass. Approximately 50% at low CO2 availability. Only weak biomass formation was
of the carbon was fermented into lactate, and 30% was con- found during the first 0–4 h under dissolved CO2 concentra-
verted into succinate. Under low CO2-HCO3- concentrations, tion of 8.74 mM. Biomass formation and succinate produc-
the ATP yield was 0.75 mol mol-1 glucose, whereas under tion enhanced in proportion as CO2 availability improved.
-
high CO2-HCO3 concentrations, the ATP yield was 2.55 and For further enhancing CO2 availability in the medium,
2.47, respectively. The ATP yield were significantly increased batch cultures were fulfi lled with varied concentrations of
-
under sufficient CO2-HCO3 conditions, suggesting that NaCO3, MgCO3, or CaCO3 implement as an additional CO2
there is a critical value of CO2 in A. succiniciproducens above source. When 119 mM of NaHCO3 or MgCO3 (both corre-
which succinate production improves significantly. The sponding 141 mM dissolved CO2 concentration) was added,
authors concluded that lactate production in A. succinicipro- biomass formation and succinate fermentation were further
ducens was controlled by high pH and succinate production increased. Compared with the yields of biomass and suc-
was controlled by CO2 availability. At pH 6.2 and sufficient cinate at dissolved CO2 concentration of 8.74 mM, the yields
CO2-HCO3- conditions, succinate accumulates as a major increased at dissolved CO2 concentration of 141 mM by 49%
product. PEP carboxykinase activity is high, but lactate and 52%, respectively. However, biomass formation and
dehydrogenase and ethanol dehydrogenase activity are lack- succinate production were inhibited to some degree in the
ing; whereas at pH 7.2 and insufficient CO2-HCO3- condi- media with 238 mM of NaHCO3 and MgCO3 (correspond-
tions, PEP carboxykinase activity is lower and lactate dehy- ing to dissolved CO2 concentration of 260 mM and 163 mM,
drogenase and alcohol dehydrogenase activity are detected. respectively). The addition of CaCO3 strongly suppressed
The effect of CO2 aeration at different pH on cell growth biomass formation because the maximum specific growth
and succinate formation were investigated by Lee et al.122 rate and the maximum dry cell weight were even less than
using A. succiniciproducens in pH-controlled batch fermen- those at dissolved CO2 concentrations of 8.74 mM.
tations at 0.25 vvm CO2 flow. Inhibited cell growth by CO2 These results tell us that CO2 availability has a great impact
aeration was observed at pH 6.2 and 6.5. At pH 6.2, there is on biomass and product formations. The pH is also a key
a constant succinate yield of 0.82–0.83 g g-1 glucose with or factor because it affects the solubility of CO2 in the medium,
without CO2 supply. However, succinate yield increased from and as a consequence influences the availability of CO2 for
0.84 g g-1 without CO2 supply to 0.88 g g-1 with CO2 supply micro-organisms. Because different micro-organisms can
at pH 6.5. At pH 7.2, both dry cell weight and succinate yield tolerate different CO2 levels during the fermentation, the
decreased sharply. They concluded that different succinate best CO2 concentration should be obtained on an individual
yields were due to different CO2 solubilities at different pH. basis for each micro-organism and medium used.
It is reasonable to explain succinate yield increase with CO2 H2
aeration at pH 6.5. However, it can’t explain why succinate In addition to CO2, H2 as a potential electron donor can
yield keeps the same at pH 6.2 with or without CO2 supply. affect cellular metabolism. Lee et al.122 found that H2/CO2
Different to succinate yield, biomass was adversely affected mixed aeration improved A. succiniciproducens biomass
by CO2 aeration. These phenomena hint that CO2 has selec- formation and succinate fermentation and that the ratio of
tive inhibition on metabolism by affecting intracellular enzy- H2/CO2 was optimized. Compared with 100% CO2 flow,
matic activities. This hypothesis was proved by the phenom- the succinate yield and productivity using 5% H2 and 95%
ena that some enzymes, particularly, involved in carboxlya- CO2 increased by 5.8% (0.86 vs 0.91 g g-1) and 80% (1 vs 1.8
tion or decarboxiation reactions can be affected by CO2. g l-1h-1), respectively. They concluded that the enhancement
The effect of the CO2 availability on biomass formation was due to the decreased cellular redox potential and the
and succinate fermentation under various CO2 partial pres- improved NADPH recycles. Therefore, the outside sup-
sures (calculated by a model developed by the authors) were plement of H2 seems to have two major functions: (i) to
investigated in batch culture using M. succiniciproducens decrease intracellular redox potential which will enhance

310 © 2012 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 6:302–318 (2012); DOI: 10.1002/bbb
Review: Biotechnological production of succinic acid K-K Cheng et al.

biomass formation and (ii) to promote glucose bioconver- succinate/acetate in pH 6.0–7.2 were stable though the con-
sion to succinate due to the incorporation of electrons centrations were different.90 The effects of some neutralizers
derived from H2. Similar phenomena were observed by Van (including MgCO3, CaCO3, Na 2CO3, NaOH and NH4OH)
der Werf et al;21 succinate production by Actinobacillus sp. were investigated for pH buffer in succinate fermentations
130Z was enhanced when 100% H2 was aerated. Compared using A. succinogenes CGMCC1593. The authors observed
with pure CO2 flow, acetate formation declined by 6% when that there was no succinate or other organic acid accumula-
external supply of 5% H2 was added. They deduced that suc- tion if NH4OH was used as a neutralization buffer, and the
cinate is a highly reduced fermentative product using four strains could not propagate due to the toxicity NH4OH.
electrons per molecule formed, and therefore the supple- When Na 2CO3 or NaOH was used, cells flocculated and
mentation of H2 may lead to the accumulation of a greater lumped after 16 h, and biomass dropped sharply. However,
amount of more reduced fermentative product succinate, cell growth was normal during the whole culture process
rather than acetate. with MgCO3 as pH neutralizer.
The supply of other electron donors, such as electrically According to Nghiem et al., 59 the optimum pH value for
reduced neutral red, also led to the obvious improvement of suc- succinate production by A. succiniciproducens ATCC 53438
cinate fermentation.66 These conclusions are in accordance with is the pH 6. At pH both above and below 6.0, the rate of cell
that the use of more reduced substrates such as mannitol and growth was significantly decreased. At pH 5.0, cell didn’t
arabitol which led to considerable enhancement in the succi- grow at all. The consumption of glucose follows the same pat-
nate yield compared with glucose (increase by 21.3% and 37.3%, tern. A range of pH 6.2–6.5 was thought to be fit for succinate
respectively), while the use of more oxided sugars such as glu- fermentation using A. succiniciproducens ATCC 29305.122
conate which resulted in decrease in the succinate yield.21,124 However, in this case, succinate formation was slightly higher
pH at higher pH (35 g l-1 at pH 6.5 compared to 33.2 g l-1 at pH
The pH is a key parameter in the bioconversion process 6.3), although the productivity (2.01 g l-1h-1 at pH 6.2 com-
because both intracellular enzymatic activities and cellu- pared to 1.92 g l-1h-1 at pH 6.5) was higher at lower pH.
lar maintenance are strictly pH dependent. Generally, the It was found that Enterococcus flavescens accumulated suc-
optimal pH for bacterial culture is about 6–7, among which cinate in range of pH 4.0–9.0. The maximum productivity of
the optimum biomass yield will be obtained. Most succinate 0.92 g l−1 h-1 was reached at pH 6.5, above which there was
production by bioconversion is accompanied by accumula- a decrease in the succinate concentration.125 The enzymes
tion of other organic acids, such as acetate and lactate. Thus, involved in succinate formation, such as phosphoenol pyru-
during succinate fermentation, if no pH buffer is included, vate carboxylase (PPC), phosphoenol pyruvate carboxyki-
the medium will acidulate gradually. The undissociated nase (PPCK), and malate dehydrogenase, showed maximum
acids, which will be harmful for biomass formation and activities at pH 6.5. Similarly it was found that M. succinicip-
substrate consumption, will increase. Due to fermentative roducens MBEL 55E showed the same cell growth at a pH of
product inhibitions, the metabolism will end. 6.0–7.5 and accumulated succinate, acetate and formate at a
With uncontrolled pH from an initial pH of 7.0, constant ratio of 2:1:1.43
Actinobacillus sp. 130Z produced acetate, formate, ethanol, Yuzbashev et al.126 developed a yeast Yarrowia lipolytica
and succinate as the major products, with a gradual pH Y-3314 in which the gene coding of one of the succinate
decline to 5.2, at which biomass formation ended. The opti- dehydrogenase subunits was deleted and could use glycerol
mal pH for biomass formation was found at 7.0. The concen- as a substrate to produce succinate even at pH below 3.5.
trations of product accumulated were almost the same in When CaCO3 was used for buffer during fermentation in
the scope of pH 6.0–7.4.21 Similar results were found in A. shaking flasks, the average succinate concentration was
succinogenes CGMCC1593; it did not propagate and did not 45.5 g l-1. When no buffer was used, the pH dropped below
accumulate succinate at pH below 5.5. The pH for the high- 3.5 after 72 h of fermentation. Nevertheless, 17 g l-1 succinate
est succinate yield was found at pH 6.7. The molar ratios of accumulation was observed in the cell stationary phase.

© 2012 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 6:302–318 (2012); DOI: 10.1002/bbb 311
K-K Cheng et al. Review: Biotechnological production of succinic acid

Integrated production with other would promote the tricarboxylic acid cycle and benefit suc-
high-value-added product cinate production. Further, polyhydroxybutyrate formation
reduced pyruvate and acetate secretion and had a positive
NADH and ATP have to be yielded for biomass formation
effect on the cell growth. Therefore, integrated production
and succinate production by oxidation of glucose or another
of succinate and polyhydroxybutyrate made the carbon flux
substrate to some products, such as acetate and lactate.
more balanced. Based on E. coli LR1110, an E. coli KNSP1
Many of these undesired products are low-value-added.
strain was developed by deletions of ptsG, sdhA and pta
Genetic engineering approaches and biorefining strategies
genes and overexpression of phaC1 from Pseudomonas aeru-
provide highly effective methods for producing desired and
ginosa. Using a pulse glycerol feeding strategy in aerobic
high-value-added compounds. Because the major object is to
cultivation, E. coli KNSP1 can produce 21.07 g l-1 succinate
produce economic and green biological succinate, a type of
and 0.54 g l-1 PHA (occupied 41.3 % of dry cell weight)
fermentation that produces two commercial interests might
from glycerol and fatty acid mixture. The generated PHA
be considered for a promising biological production process.
composed 58.7 mol% 3-hydroxyoctanoate and 41.3 mol%
Using a low level of alkali metals medium (total salts 4.2 g
-1
l ), E. coli KJ073(ldhA, adhE, ackA, focA, pflB, mgsA, poxB) 3-hydroxydecanoate. This strain would be useful for utiliza-
produces 78.9 g l-1 succinate and 15.8 g l-1 malic acid in a tion of by-product glycerol and fatty acid from the biodiesel
batch fermentation with initial 100 g l-1 glucose, strain KJ071 production process.128
(ldhA, adhE, ackA, focA, pflB, mgsA), can produce 33.1 g l-1 Glycerol metabolism in Klebsiella pneumoniae includes
succinate and 69.2 g l-1 malic acid with total molar yields two routes: the reductive route and the oxidative route. In
(succinate+malate) of 2.2 per mole of glucose metabolized. the reductive route, glycerol is first converted to 3-hydroxy-
This study could have the advantages of flexible produc- propanal by glycerol dehydratase. 3-Hydroxypropanal is
tion using different microorganisms (KJ073 or KJ071) and then converted to 1,3-propanediol by 1,3-propanediol oxi-
producing different compounds (succinate or malic acid) doreductase. In the oxidative route, some by-products, such
depending on the market situation.18 as 2,3-butanediol, acetate, lactate, succinate, and ethanol, are
An isoamyl acetate production pathway was designed produced. K. pneumoniae HR526, a new screened organism,
in E. coli by expression pyruvate carboxylase and alcohol exhibited high productivity but excrete lactate in the late-
acetyltransferase for accumulation isoamyl acetate and suc- exponential phase as the main by-product. When K. pneu-
cinate with high yields of both high value products.127 Since moniae LDH526 was developed with D-lactate pathway dele-
the two products have different volatility, they can be easily tion, intra-cellular NADH/NAD+ increased significantly.
purified. Using 20 g l-1 glucose and 0.8 g l-1 isoamyl alcohol, Due to more NADH formation in the late-exponential
E. coli SBS990MG(pHL413, pKmAT) produced 1.22 g l-1 phase, cell growth in fed-batch fermentation by K. pneumo-
isoamyl acetate and 5.37 g l-1 succinate. In this process, suc- niae LDH526 was quicker and biomass increased by about
cinate formation was used to keep the reasonable balance 6% at the stationary phase. 1,3-propanediol concentration
between NADH and NAD+ to maximize isoamyl acetate increased from 95.4 to 102.1 g l-1. Succinate formation, as an
yield. alterative NADH consumption route, was also enhanced and
A genetically modified E. coli QZ1112 was constructed its concentration increased from 9.2 to 13.8 g l-1.129
by inactivation of sdhA, Pta, and PoxB gene and express- A biorefining strategy based on solid state fermentation
ing phbCAB genes from Ralstonia eutropha to accumulate using wheat as feedstock for succinate production was
extra-cellular succinate and intra-cellular polyhydroxy- developed by Du et al.118 At first, wheat was fractionat-
butyrate at the same time in aerobic culture.22 In a batch ingly treated to bran, gluten, and gluten-free flour by size
culture using 45 g l-1 glucose, E. coli QZ1112 can accumulate reduction and extraction. The bran was used for Aspergillus
24.6 g l-1 succinate and 4.95 g l-1 polyhydroxybutyrate, which awamori glucoamylase preparation and Aspergillus oryzae
occupied 41.3% of dry cell weight. During polyhydroxybu- protease preparation by solid state fermentation, respec-
tyrate formation, a great deal of NADPH was used, which tively. The prepared raw enzymes were separately utilized

312 © 2012 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 6:302–318 (2012); DOI: 10.1002/bbb
Review: Biotechnological production of succinic acid K-K Cheng et al.

to hydrolyze flour and gluten to produce over 140 g l-1 of expresses a cellulolytic or a cellulolytic enzyme gene or
glucose solution and a more than 3.5 g l-1 of amino nitrogen engineering cellulolytic organism with succinate production
solution. A mixed medium consisting of these two solutions properties. By studying metabolic regulation and metabolic
contained all the necessary nutrients required for succinate engineering bacteria, combined with physical and chemi-
production. In a fermentation using A. succinogenes ATCC cal pre-treatment technology, it is expected to design new
55618 from the mixed medium and addition of MgCO3, integrated biorefinery technology to transform lignocellu-
around 64 g l-1 succinate was obtained. These results dem- losic biomass into succinate successfully.
onstrate that wheat can be efficiently utilized for the produc- Recovery and purification of succinate represents a techno-
tion of succinate and other high-value-added products by logical challenge and an economical obstacle for an efficient
biorefining strategies. microbial production on a large scale. All methods used at
present showed unsatisfied product yield and purity. For fur-
Conclusions and perspectives ther development, traditional separation techniques should to
Succinate has been included in the US Department of be improved and coupled with upstream technology. For exam-
Energy’s Top Value Added Chemicals from Biomass based ple, by target deletion of acid by-products using genetic modi-
on its potential to become an important building block for fication to lower or eliminate acid by-products’ accumulation,
deriving both commodity and specialty chemicals.130 Three the selectivity of succinate in reactive extraction over other
important process parameters decide the economical viabil- acid by-products will be improved. Since product inhibition is
ity of a bioprocess: yield, concentration, and productivity. found in succinic acid fermentation,131 another approach worth
The yield relates more to the variable cost of raw feedstock considering and an attractive method for increasing productiv-
and will be of increasing importance with increasing sugar ity of the process is fermentation combined with in situ product
prices. Productivity and concentration relate more to the removal. Such an approach, if precisely optimized, could result
fi xed cost and total investment. Low rates imply larger in a highly efficient bioprocess. Furthermore, a novel separation
energy and labor cost; low concentrations will result in method using an emulsion liquid membrane, which shows high
larger energy input for product recovery. Compared with selectivity in the separation of acetate from succinate, deserves
petrochemical-derived succinate, biological succinate pro- attention in the future.132
duction is still not economically competitive, because of its Biological succinate can decrease the use of non-renewable
major drawbacks: high cost of the feedstocks, low product resource and reduce greenhouse gas emissions. Currently,
concentration in the fermentation broth, the co-production the demand for succinate has been entering a time of rapid
of low-value acid by-products, and difficult product recovery. increase, bringing both profits and a driving force. We
To increase the competitiveness of the biological produc- believe that through extensive industry-university-research
tion of succinate, future work should be addressed as fol- co-operation, a sustainable and economically viable process
lows: increasing succinate concentration and yield through for biosuccinate production can be established in the near
metabolic engineering; introducing and optimizing the suc- future.
cinate synthetic pathway in the species with high succinate
Acknowledgements
tolerance; overcoming the substrate repression effect and
This study was supported by National Natural Science
utilizing low-cost non-food-based feedstocks; and integrat-
Foundation of China (21176139) and National Basic Research
ing production of succinate with other high-value-added
Program of China (2011CB707406).
products. Lignocellulose is the most abundant renewable
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K-K Cheng et al. Review: Biotechnological production of succinic acid

Keke Cheng Jing Zeng

Dr Keke Cheng received his PhD from Jing Zeng received her Master’s degree from
the Department of Chemical Engineering, the Department of Chemical Engineering,
Tsinghua University in 2005. He has worked Tsinghua University in 2006. She has worked
at the Institute of Nuclear and New Energy at the Department of Chemical Engineering,
Technology, Tsinghua University, as an Tsinghua University, as a research assist-
Assistant Professor since April 2005. His ant since April 2006. Her specialized field is
specialized fields are biorefinery, biofuel and biofuel policy.
fermentation technology.

Xuebing Zhao Jian-An Zhang

Dr Zhao obtained his PhD in 2009 and Dr Jian-An Zhang is Associate Professor
continued his post-doctoral work at the at the Institute of Nuclear and New Energy
Department of Chemical Engineering, Technology in Tsinghua University. His main
Tsinghua University. He became an Assistant research interests are bioenergy (bioethanol,
Professor in the same department in 2011. biodiesel, biobutanol), biochemical engi-
His current research topics are the biorefining neering and the utilization of lignocellulose
of lignocellulosic biomass to produce biofuels biomass.
and chemicals.

318 © 2012 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 6:302–318 (2012); DOI: 10.1002/bbb