Separation and Purification Technology 72 (2010) 294–300

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Separation and Purification Technology

journal homepage: www.elsevier.com/locate/seppur

One step recovery of succinic acid from fermentation broths by crystallization

Qiang Li a,b , Dan Wang a,b , Yong Wu b,c , Wangliang Li a , Yunjian Zhang d , Jianmin Xing a,∗ , Zhiguo Su a
a
National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, P.O. Box 353, Beijing 100190, PR China
b
Graduate School of the Chinese Academy of Sciences, Beijing 100049, PR China
c
Key Laboratory of Green Process and Engineering, Institute of Process Engineering, Chinese Academy of Sciences, P.O. Box 353, Beijing 100190, PR China
d
Department of Bioengineering, School of Chemical and Environmental Engineering, China University of Mining and Technology (Beijing), Beijing 100083, PR China

a r t i c l e i n f o a b s t r a c t

Article history: Succinic acid is a valuable four-carbon platform chemical with wide applications in many fields. One
Received 19 July 2009 step recovery of the desired product succinic acid crystal from the fermentation broths was studied for
Received in revised form 21 February 2010 the first time. In a fed batch fermentation by Actinobacillus succinogenes BE-1, the final concentrations of
Accepted 24 February 2010
succinic acid, formic acid, lactic acid and acetic acid in the bioreactor reached 97.8 g/L, 23.5 g/L, 5.1 g/L
and 17.4 g/L, respectively. These organic acids have different proportions of dissociated and undissociated
Keywords:
forms at different pH values, and the solubility of these forms of acid is different substantially. When the
Crystallization
pH value of the fermentation broth was controlled less than 2.0, crystallization of succinic acid could
Succinic acid
pH be carried out at 4 ◦ C easily, while major acid by-products such as lactic acid, acetic acid and formic
Temperature acids were miscible in solution. By this one step recovery method, succinic acid yield was 70%, and the
purity was 90%. In comparison, for traditional calcium precipitation coupled ion-exchange adsorption,
the yield and purity were 52% and 92%, respectively. Crystallization could be regarded as not only the final
purification step but also the first recovery step for the downstream separation process. In this study,
succinic acid can be feasibly purified from fermentation broths selectively, which could be integrated
with other separation method to optimize the downstream process.
© 2010 Elsevier B.V. All rights reserved.

1. Introduction Ca(OH)2 is usually added to neutralize the fermentation broth and

Succinic acid is considered as one of the twelve building
block organic compounds and has drawn worldwide concerns for
its industrial applications in biodegradable polymers, foods, fine
chemicals and pharmaceuticals [1–3]. Traditional petrochemical
production of succinic acid requires high temperature, high pres-
sure and costly catalysts. Therefore, much attention has been paid
to the microbial conversion of biomass to succinic acid and the
recovery of succinic acid from the fermentation broth recently [4].
Production of succinic acid at high titers and high yield has been
widely reported. For example, the final fermentation titers of suc-
cinic acid can reach over 50 g/L by Mannheimia succiniciproducens
[5], 83 g/L by Anaerobiospirillum succiniciproducens [6], 99.2 g/L by
E. coli [7] or even higher than 100 g/L by Actinobacillus succinogenes
[8,9].
Downstream recovery process is estimated to account for over
60% of the overall production cost. The existing separation meth-
ods are usually not easily operated to separate target acid from the
carboxylic acid mixture in the fermentation broths. Traditionally,
∗ Corresponding author. Tel.: +86 010 62550913; fax: +86 010 62550913.
E-mail addresses: qiangli@home.ipe.ac.cn (Q. Li), jmxing@home.ipe.ac.cn
(J. Xing).
precipitate succinate [8,10]. Calcium succinate is recovered by fil-
tration, and free acid is released from the precipitate by adding
sulfuric acid. However, large amount of CaSO4 and some Calcium
lactate could be accumulated. The reactive extraction with amine-
based extractant has been reported in many literatures [11–15].
This method is easy to operate by controlling the pH value of the
two phases. Electrodialysis is used to recover succinate from non-
ionized compounds with proper ion exchange membrane [6,16].
Adsorption has been widely used in many organic acids separa-
tion since adsorbents have advantages of quick recovery, and low
regeneration consumption [17,18]. A vacuum distillation method
was developed to eliminate by-product volatile carboxylic acids,
such as acetic, formic acids from the broths at pH 4.2, 60 ◦ C. Then,
succinic acid was crystallized from the mother solution [19]. Many
processes were often interrupted by the contaminated acids, impu-
rities, carbon sources, protein, or salts, with crystallization process
as the final purification step to refine pure succinic acid [20].
The recovery processes are still not economically feasible
because the properties of the carboxylic acids in the acid mixture
are similar. According to the physicochemical characteristics, these
short chain organic acids have the similar behavior in solution at
room temperature. As for succinic acid, pKa1 is 4.21, and pKa2 is
5.64. For lactic acid, pKa is 3.86. pKa of acetic acid is 4.7 and pKa
of formic acid is 3.84 [21]. Thus, in many fermentation processes,

1383-5866/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.seppur.2010.02.021
 Q. Li et al. / Separation and Purification Technology 72 (2010) 294–300
the mixture acids were assumed to be the sum of the individual
acids to simplify the process modeling [21,22]. However, it is not
simple to remove the target acid from the mixture acids effectively
in the downstream process because many separation methods do
not have the perfect selectivity toward succinic acid out from the
other carboxylic acids.
In this paper, the possibility of one step recovery of succinic acid
from the mixture acids in the fermentation broth directly by crys-
tallization was studied theoretically and experimentally. Moreover,
recovery effectiveness of the new separation process was com-
pared with the calcium precipitation process coupled an adsorption
operation.
2. Materials and methods
2.1. Microorganisms, chemicals and fermentation
A. succinogenes BE-1, a strain that produced a high concentra-
tion of succinic acid, had been isolated from bovine rumen in our
laboratory, and was collected in China General Microbiological Cul-
ture Collection Center (No. CGMCC2650). All chemicals used in
this study were of analytical level and were purchased from either
OXOID (England) or Sinopharm Chemical Reagent Beijing Co., Ltd.
(China) unless otherwise described.
Effect of different carbon sources on the production of succinic
acid was carried out in 1 L flask fermentation. The flask fermen-
tation medium contained per liter: 30 g sugar, 30 g yeast extract,
2 g urea, 2 g MgCl2 ·6H2 O, 1.5 g CaCl2 , 0.07 g MnCl2 , 4.4 g Na2 HPO4 ,
3.3 g NaH2 PO4 , 30 g MgCO3 . Effect of pH neutralization agents on
the production of succinic acid was carried out in batch fermen-
tation in a 5 L NBS stirred bioreactors (New Brunswick Scientific
Co., Inc. USA) containing 3 L of medium. pH was strictly maintained
at 6.8 with 10 mol L−1 NaOH, NH3 OH, 3 mol L−1 Na2 CO3 , 50 g L−1
solid MgCO3 as the pH buffer. The fermentation medium contained
per liter: 60 g glucose, 30 g yeast extract, 2 g urea, 2 g MgCl2 ·6H2 O,
1.5 g CaCl2 , 0.07 g MnCl2 , 4.4 g Na2 HPO4 , 3.3 g NaH2 PO4 . The seed
medium contained per liter: pancreatic digest of casein 17 g L−1 ,
soy peptone 3 g L−1 , glucose 2 g L−1 , NaCl 5 g L−1 , K2 HPO4 2.5 g L−1 .
Seed cultures were prepared at 37 ◦ C, 160 rpm in flask. Inocula-
tion volume was 5% (v/v). Sugar was separately sterilized at 115 ◦ C
for 30 min and fed-batch added to the medium. The initial pH of
the sterilized medium was adjusted to 7.0 by 10 mol L−1 of NaOH
solution. Fed batch fermentation was carried out in a 5 L NBS
bioreactor containing 3 L of medium. The fed-batch fermentation
medium contained per liter: 60 g glucose, 30 g yeast extract, 2 g
urea, 2 g MgCl2 ·6H2 O, 1.5 g CaCl2 , 0.07 g MnCl2 , 4.4 g Na2 HPO4 , 3.3 g
NaH2 PO4 , 80 g MgCO3 . The temperature and stirring speed were
regulated at 37 ◦ C, 150 rpm. CO2 was continuously sparged in the
reactor at the rate of 1.0 vvm.
2.2. Recover process
Cells were separated by centrifugation for 20 min at 8000 rpm.
Activated carbon (20%, w/v) was mixed with the filtrated fermen-
tation broth overnight to decolour the broth and remove glucose
residuals or protein impurities. The suspension was filtered and
then treated using either direct crystallization method (Process I)
or modified calcium precipitation method (Process II) to recover
succinic acid. The schematic diagram was shown in Fig. 1.
In Process I, the pH of the aqueous broth was adjusted to 2.0 by
the addition of 35% (v/v) HCl. The acidification rate was 1 pH value
per min. Then, the clear filtrated fermentation broth was cooled to
2–4 ◦ C, and the succinic acid was crystallized to a constant weight in
a 2 L crystallizer with 90 rpm agitation rate for at least 5 h. Crystals
were gathered by centrifugation for 10 min at 8000 rpm, 4 ◦ C.
295
In Process II, Ca(OH)2 slurry (25–33%, w/v) was slowly added
to the clear filtrated fermentation broth with a stirring mixture at
300 rpm at room temperature until the pH rose up to 12.5. Solid
calcium succinate was washed with distilled water several times.
Free acid was released from calcium succinate by adding H2 SO4
(40–60%, w/v). The addition of sulfuric acid was stopped when the
pH decreased to around 2.0 and the precipitate of calcium sulfate
was removed by filtration. The ion-exchange adsorption of succinic
acid in the filtrate was carried out in a similar way [18]. The filtrate
was fed to the anion exchange column with the flow rate 1 mL/min.
0.5 mol L−1 of HCl was chosen as an elution agent to recover suc-
cinic acid from the adsorbents. The elute solution was dried at 60 ◦ C
overnight to obtain succinic acid crystals.
2.3. Analytical methods
For determination of biomass, 5 mL of samples were removed
from the culture, centrifuged, washed once with 1 M HCl and twice
with distilled water, finally dried to a constant weight at 60 ◦ C.
Concentrations of glucose were measured by SBA 40 C bio-sense
analyzer (Biology Institute, Shandong Academy of Sciences, China).
The different reductive sugar was estimated by the traditional DNS
method. The concentrations of organic acids were measured by
high-performance liquid chromatography (Agilent, USA) equipped
with an Agilent model DAD UV-VIS detector and the ZORBAX SB-Aq
column (250 mm × 4.6 mm, Agilent). The column was operated at
25 ◦ C. The mobile phase was pH 2.7, 20 mmol L−1 KH2 PO4 at a flow
rate of 1 mL min−1 . The injection volume was 10 ␮L.
The distribution diagram of carboxylic acid products, both in
their free acid forms and their carboxylate forms, was calculated by
the followed equations according to their dissociation constants, for
example, acetic acid (Ka = 6.2 × 10−5 ), formic acid (Ka = 2.1 × 10−4 ),
lactic acid (Ka = 1.37 × 10−4 ) and succinic acid (Ka1 = 2.1 × 10−4 ,
Ka2 = 2.3 × 10−6 ). For the monoacid, distribution diagram was cal-
culated as follows:
[HA] 1 1 [H+ ]
ıHA = − = − = + = +
[HA] + [A ] 1 + [A ]/[HA] 1 + Ka /[H ] [H ] + Ka
(1)
[A− ] Ka
ıA− = = (2)
[HA] + [A− ] [H+ ] + Ka
For the diacid, distribution diagram was calculated as follows:
1 [H+ ]
2
ıH2 A = =
+ 2 + 2
1 + Ka1 /[H+ ] + Ka1 Ka2 /[H ] [H ] + Ka1 [H+ ] + Ka1 Ka2
(3)
Ka1 [H+ ]
ıHA− = 2
(4)
[H+ ] + Ka1 [H+ ] + Ka1 Ka2
ıHA− = 1 − ıHA− − ıH2 A (5)
where ı was the distribution (%) of the free acid forms/ionic forms,
Ka was dissociation constant, and [H+ ] was the concentration of the
H+ which controlled by the pH value of the solution.
The succinic acid solubility measurements were easily done
under normal pressure and at the temperature 1–80 ◦ C. The solubil-
ity of succinic acid equals the concentrations of saturated succinic
acid solution. The pH of the mother solution was controlled by
HCl and NaOH. The concentration of succinic acid solution and
the recovery yield were measured by HPLC. Scanning electron
microscopy (SEM) was used for morphological analysis. Yield and
purity were calculated as followed, yield (%) = (dry weight of suc-
cinic acid in crystals recovered (g)/weight of succinic acid in the
initial fermentation broth (g)) × 100%, purity (%) = (dry weight of
296 Q. Li et al. / Separation and Purification Technology 72 (2010) 294–300

Fig. 1. Schematic diagrams of two recovery processes of succinic acid, (I) direct crystallization, (II) modified calcium precipitation.

succinic acid in crystals recovered (g)/dry weight of crystals recov-
ered (g)) × 100%.
3. Results
3.1. Fermentation of A. succinogenes BE-1
Effect of different carbon sources on the production of succinic
acid was carried out in flask fermentation. As shown in Table 1
batches 1–5, A. succinogenes could utilize glucose, fructose, xylose,
maltose and cellobiotose to produce succinic acid. For xyltose as
the carbon source, cell growth and fermentation were inhibited.
For cellobiotose as the carbon source and the sugar utilization
was lower than 50%. The succinic acid conversion yield on glucose
was relatively higher than other sugars. Thus, glucose was used as
the carbon source for batches 6–10 while the neutralization agent
was varied. Effect of pH neutralization agents on the production
of succinic acid was carried out in stirred bioreactors with an ini-
tial glucose concentration of 60 g L−1 . pH was strictly maintained
at 6.8 with 10 mol L−1 NaOH, NH3 OH, 3 mol L−1 Na2 CO3 , 50 g L−1
solid MgCO3 . As shown in Table 1, MgCO3 could act as one of the
best neutralizers which help to produce succinic acid at high titers
and yield. With the comparison of the batches 6 and 7, it can be
found CO2 was useful to increase the titers and yield of the prod-
uct. Thus, an optimized fed-batch fermentation of A. succinogenes
could be carried out with MgCO3 as the pH buffer and continu-
ous CO2 sparging. A. succinogens BE-1, a gram-negative bacterium,
produced a substantial amount of succinic acid as a major fermen-
tation product under anaerobic conditions. Fig. 2 showed a time
profile of metabolite production and the glucose consumption dur-
ing an optimized fermentation of A. succinogenes BE-1 with batch
glucose and yeast extract feeding at 8 h, 15 h and 24 h. Like other
 Q. Li et al. / Separation and Purification Technology 72 (2010) 294–300 297

Table 1
Effect of carbon sources and pH neutralization agents on the production of succinic acid in batch fermentation.

Batch Carbon source Titers (g L−1 ) Yield (g L−1 h−1 ) Batch Neutralization agent Titers (g L−1 ) Yield (g L−1 h−1 )

1 Glucose 19.2 ± 0.1 0.51 ± 0.06 6 NaOH 8.9 ± 0.2 0.24 ± 0.04
2 Fructose 13.1 ± 0.3 0.42 ± 0.05 7 NaOH + CO2 29.1 ± 0.1 1.12 ± 0.08
3 Xyltose 5.3 ± 0.2 0.21 ± 0.04 8 NH3 OH + CO2 15.5 ± 0.3 0.89 ± 0.11
4 Maltose 14.8 ± 0.1 0.50 ± 0.03 9 Na2 CO3 + CO2 39.2 ± 0.1 1.24 ± 0.08
5 Cellobiotose 15.1 ± 0.4 0.43 ± 0.05 10 MgCO3 + CO2 53.3 ± 0.1 2.43 ± 0.05

3.2. Proportions of dissociated and undissociated carboxylic acid

at different pH values

Succinic, lactic, formic and acetic acids have different propor-

tions of dissociated and undissociated forms at different pH values.
The distribution diagram ı (%) reflected proportions of the car-
boxylic acids in their free acid form or salt forms. As shown in
Fig. 3, the ı curves represented the accurate forms of these acid
products. For formic, lactic and acetic acid, their 90% ionic forms
were obtained when the pH value of the solution was 6.0. If the pH
value of the solution was above 7.0, all the three acids were in their
ionic forms. However, succinic acid, as a dicarboxylic acid, was dif-
ferent. Only when the pH value of the aqueous solution was above
8.0, 100% succinic acid was in its ionic form. If the pH value was
adjusted less than 2.0, all these carboxylic acids were in their free
acid forms.
Fig. 2. Time profile of metabolite production and the glucose consumption during
the fed-batch fermentation with A. succinogenes BE-1.
3.3. Solubility curves of succinic acid and succinate

succinic acid-producers, A. succinogens BE-1 also produced acetic,
formic, and lactic acids as the major by-products. At 35 h, the titers
of succinic acid were 97.8 g/L, with 75% glucose conversion yield.
Final concentrations of formic, lactic and acetic acid were 23.5 g/L,
5.1 g/L and 17.4 g/L, respectively.
Lactic, formic and acetic acids, no matter in their free acid forms
or salt forms, are miscible within water at different pH values.
However, the solubility of succinic acid changed substantially at
different pH values as a result of dissociated forms. pH can eas-
ily exchange between succinic acid and succinate, while succinic
acid and succinate have dramatically different solubility at varied

Fig. 3. Distribution diagram of formic/formate (A), lactic/lactate (B), acetic/acetate (C), and sucininic/succinate (D).
298 Q. Li et al. / Separation and Purification Technology 72 (2010) 294–300

Fig. 4. Solubility curve of succinic acid at 25 ◦ C at different pH controlled by HCl and

NaOH.

temperature. As shown in Fig. 4, pH had an effect on the soluble

behavior of succinic acid at the room temperature. At pH 4.0, there
were 61% succinic acid, 38% succinate− and 1% succinate2− in water
(Fig. 3D). It would be nearly 400 g/L overall succinic acid that could
be fully miscible in water. When the pH value decreased to less than
2.0, succinic acid was all in its free acid form with the solubility of
73 g/L.
Formic, acetic and lactic acids, whatever their forms are free
acid or carboxylate, have perfect solubility within aqueous broth at
pH 1.0–14 at temperature above 0 ◦ C. As shown in Fig. 5, succinate
was much more miscible in water than succinic acid. Although the
solubility of succinic acid, monosodium succiniate and disodium
succinate decreased sharply when the temperature decreased, suc-
cinic acid was much less miscible than succinates. According to
Fig. 3D, if pH was adjusted less than 2.0, succinate was totally
changed into its free acid form. At 4 ◦ C, the solubility of succinate
was nearly 200 g/L, while the solubility of succinic acid was only
30 g/L.

3.4. Succinic acid crystallization from the fermentation broth

Normally succinic acid fermentation is operated at pH 6–7,

above pKa of each acid. The final acid products are almost in their
salt forms rather than free acid. The solubility of succinic acid was
only 3% at 4 ◦ C, at pH 2.0. While the other acid by-products were
still water miscible. Therefore, when the pH was adjusted to 2.0
with HCl, succinic acid could be selectively crystallized. In this cir-
cumstance, crystallization could be regarded as a powerful tool to Fig. 5. Solubility curve of succinic acid and succinate at different temperatures.

separate the target acids from the broth. By the method of the novel
recovery approach (Fig. 1 Process I), 70% succinic acid yield and 90%
purity were obtained. In comparison, by the method of traditional
calcium precipitation coupled ion-exchange adsorption (Fig. 1 Pro-
cess II), the yield and purity were only 52% and 92%, respectively.
Fig. 6 showed the SEM image (300×) of succinic acid after each
purification process. Fig. 6A was the succinic acid image of com-
mercial analytical reagent. Fig. 6B was an image of succinic acid
crystallized directly from the fermented broth (Fig. 1I), the mor-
phology of which was similar to that of commercial reagent, with
few host cells and proteins near the surface of the crystal. In Fig. 1
process II, Ca(OH)2 was added to precipitate the succinate. Precip-
itation of calcium succinate was recovered by filtration, and free
acid was released from the precipitate by adding sulfuric acid. The
precipitate of calcium sulfate was removed by filtration. The fil-
trate could be also crystallized to succinic acid crystal (Fig. 6C), or
adsorbed by an anion exchange column, regenerated and dried to
succinic acid crystals (Fig. 6D). As shown in Fig. 7, the IR image
showed the purity of the purified product according to process I
(Fig. 6B) and process II (Fig. 6D). The main bands appeared in the
500–3500 cm−1 range. Compared with the standard IR image of
the succinic acid, the purity of the final purified succinic acid was
very much close to the standard image [23] although there was
morphological difference between SEM images.
4. Discussion
For the production of organic acids, the fermentation would not
be run at low pH, most preferably with requiring neutralization. The
cost of neutralization is reasonable and acceptable, but the conver-
sion of the salt into the free acid increases cost significantly [3]. The
solubility of succinate− or succinate2− varies dramatically at the
different pH. Normally when the pH of the fermentation process
was controlled at 6.8, the succinic acid solutions contained 93.5%
succinate2− , 6.4% succinate1− and 0.1% free acid (Fig. 3). At pH 2.0,
 Q. Li et al. / Separation and Purification Technology 72 (2010) 294–300
Fig. 6. SEM image succinic acid: (A) analytical reagent of succinic acid; (B) succinic acid crystals by process I; (C) succinic acid crystals by process II (without ion exchange
separation); (D) succinic acid crystals by process II.
succinic acid solution contained 0.6% succinate1− and 99.4% free
acid. By controlling the low pH and temperature the target acid
could be crystallized, while the other acid by-products were still
fully miscible in water in their free acid forms or salt forms. Thus,
direct crystallization (Process I) is a powerful method to recover
the free acid in a downstream separation process, which could
be integrated with any other effective separation methods. The
methodology used in this study could be useful especially for the
optimization of succinic acid downstream processes.
Regardless of the separation methods, recovering succinic acid
is a significant expense. Thus, using an easy separation method and
an optimized fermentation procedure that produces succinate at
high titers will contribute to cost-effectiveness. Moreover, fermen-
tation process could have a significant influence on the downstream
recovery. For example, high product titers in the final fermentation
Fig. 7. IR image of succinic acid.
299
broth could be very helpful for the crystallization process as a result
of solubility distances. Meynial-Salles et al. reported that the final
concentrations of succinate were 83 g/L in an integrated membrane
bioreactor process [6]. By calculation theoretically, the recovery
acid yield of one step crystallization may reach 64%, which means
the cost of down stream separation can be cut dramatically. The
36% residuals in the mother liquor could be treated following addi-
tional separation methods. Adsorption process seemed efficient for
succinic acid recovery at low titers. Process II is a traditional cal-
cium precipitation method coupled ion-exchange adsorption. Like
other calcium precipitation process [19], the yield was relatively
low, only 52%, and commercially unusable gypsum was formed
inevitably. However, In Process I, the purity (90%) was lower than
Process II (92%), which was due to the relatively fast acidification
rate and the fast cooling rate. If the crystallizing was too fast, a
high supersaturation maybe created and impurities can be easily
trapped.
In the study, about 70% succinic acid can be recovered from
the fermentation broth directly by the one step crystallization.
The residual mother liquor (without the crystals) can be purified
through conventional scheme (Process II), or vacuum distillation
in a similar way to that reported by Luque et al. [19]. Vacuum
distillation method was operated at 60 ◦ C for 12 h to eliminate
by-product volatile carboxylic acids at pH 4.2, and concentrate suc-
cinic acid titers in the liquor. When the solution was concentrated
to around 20% of its original volume, the crystallization of suc-
cinic acid was carried out at 4 ◦ C, which allowed the isolation of
highly pure crystals in a similar way to that reported by Song et al.
[11,14].
The succinic acid crystals can be characterized by XRD or ATR-
FTIR to determine optimal cooling curves for batch crystallization
of succinic acid [23]. Fig. 6 showed the different SEM images of
succinic acid after each purification process. In Process I the main
impurity in the crystals obtained was host cell residuals and pro-
300 Q. Li et al. / Separation and Purification Technology 72 (2010) 294–300
teins, which requiring further refine treatment for obtaining the
reagent grade succinic acid. Fig. 6D was different from the oth-
ers, which might due to the effects of crystallization temperature
[23,24] or the other carboxylic acids [25]. The other carboxylic acids
in the mother liquid could have effects on the crystallization pro-
cess because carboxylic acids have stronger affinity toward the
surface of crystal particles. Nevertheless, crystallization is one of
the oldest yet the most effective process for the preparation of solid
products. High purities and stabilities can be obtained with distinct
morphologies and functional properties. In addition, crystallization
coupled other separation methods will obviously increase the final
yield of the total recovery processes.
5. Conclusions
Direct recovery of succinic acid from A. succinogenes was set
up by the reducing the broth pH to 2 and temperature to 4 ◦ C at
the end of fed batch fermentation, taking advantage of the rela-
tively low solubility of the undissociated form of the succinic acid
compared to other small organic acid fermentation by-products.
As one of the oldest but most effective processes for the prepara-
tion of succinic acid crystals, crystallization might directly provide
the desired product (in solid or crystal form) without the need for
auxiliary components in the product-removing phase. Crystalliza-
tion process could be regarded as not only the final refine step but
also the first recovery step for the downstream separation process,
which would be an efficient and economic supplement method for
the downstream bioprocessing of the platform chemical succinic
acid.
Acknowledgements
This work was supported by the Knowledge Innovation Program
of the Chinese Academy of Sciences (Grant No. KSCX2-YW-G-021),
and State Major Basic Research Development Program of China
(Grant 2007CB714305).
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