Bioresource Technology 135 (2013) 396402

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Two-stage in situ gas stripping for enhanced butanol fermentation

and energy-saving product recovery
Chuang Xue a,b, Jingbo Zhao b, Fangfang Liu b, Congcong Lu b, Shang-Tian Yang b,, Feng-Wu Bai a
a
b

Department of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China
Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, USA

h i g h l i g h t s
" High-titer n-butanol was produced by C. acetobutylicum in a brous bed bioreactor.
" In situ gas stripping for butanol separation alleviated butanol toxicity in the fermentation.
" Two-stage gas stripping produced butanol at a high titer of >400 g/L (500 g/L ABE).
" This process can produce n-butanol with greatly reduced energy consumption.

a r t i c l e

i n f o

Article history:
Available online 25 July 2012
Keywords:
ABE fermentation
Butanol
Clostridium acetobutylicum
Fibrous bed bioreactor
Two-stage gas stripping

a b s t r a c t
Two-stage gas stripping for butanol recovery from acetone-butanol-ethanol (ABE) fermentation with
Clostridium acetobutylicum JB200 in a brous bed bioreactor was studied. Compared to fermentation
without in situ gas stripping, more ABE (10.0 g/L acetone, 19.2 g/L butanol, 1.7 g/L ethanol vs. 7.9 g/L acetone, 16.2 g/L butanol, 1.4 g/L ethanol) were produced, with a higher butanol yield (0.25 g/g vs. 0.20 g/g)
and productivity (0.40 g/L
h vs. 0.30 g/L
h) due to reduced butanol inhibition. The rst-stage gas stripping
produced a condensate containing 175.6 g/L butanol (227.0 g/L ABE), which after phase separation
formed an organic phase containing 612.3 g/L butanol (660.7 g/L ABE) and an aqueous phase containing
101.3 g/L butanol (153.2 g/L ABE). After second-stage gas stripping, a highly concentrated product containing 420.3 g/L butanol (532.3 g/L ABE) was obtained. The process is thus effective in producing
high-titer butanol that can be puried with much less energy.
2012 Elsevier Ltd. All rights reserved.

1. Introduction
Butanol is an important chemical and advanced biofuel, which
can be produced through acetonebutanolethanol (ABE) fermentation by various Clostridium spp. (Qureshi et al., 2008). As a substitute for petroleum fuel, butanol is superior to ethanol due to its
higher energy density, less corrosiveness to existing infrastructure
and better compatibility with gasoline (Gheshlaghi et al., 2009;
Drre, 2007). However, one of the biggest challenges for biobutanol production is intensive energy consumption in product recovery from the dilute fermentation broth and low butanol yield and
productivity due to the co-production of acetone and ethanol as
well as severe butanol toxicity to cells (Ezeji et al., 2010; Nicolaou
et al., 2010; Qureshi and Blaschek, 2001b).
Metabolic engineering and mutagenesis of Clostridia have been
explored to provide solutions for enhancing butanol tolerance and
productivity (Huang et al., 2010; Papoutsakis, 2008; Zheng et al.,
Corresponding author. Tel.: +1 614 292 6611; fax: +1 614 292 3769.
E-mail address: yang.15@osu.edu (S.T. Yang).
0960-8524/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.biortech.2012.07.062

2009). However, compared to 1215 ethanol achieved in fermentation with yeasts (Xue et al., 2010), the 1.22.0 butanol produced by
solventogenic Clostridia in ABE fermentation is much lower and
not economically competitive for butanol recovery by conventional
distillation (Chen and Blaschek, 1999). Therefore, in situ butanol removal from the fermentation system by adsorption (Nielsen and
Prather, 2009; Qureshi et al., 2005), liquidliquid extraction (Rofer et al., 1987; Rofer et al., 1988), pervaporation (Matsumura
et al., 1988; Qureshi and Blaschek, 1999) and gas stripping (Qureshi and Blaschek, 2001a; Qureshi et al., 2008) has been used to mitigate butanol inhibition and improve productivity. However, a
common problem of these techniques is their low efciency in
recovering butanol with high concentration even though the productivity of the fermentation system could be improved
effectively.
Among butanol recovery methods, gas stripping has the advantages of simple operation, no harm to culture, and low energy input
and capital investment for facilities (Evans and Wang, 1988; Ezeji
et al., 2007a; Garcia et al., 1986; Qureshi and Blaschek, 2001a;
Zheng et al., 2009). However, the solvent concentration obtained

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C. Xue et al. / Bioresource Technology 135 (2013) 396402

Table 1
ABE fermentations with gas stripping.
Process

Strain

Butanol in fermentor with/without

gas stripping g/L

Condensate
Butanol g/L

ABE g/L

Batch

C. acetobutylicum P262

<5/8.1
27.2/5.2
1.98.8/11.9
26/7.4
1.33.2/11.8
3.514.6/16.2

1743
35.9
175.6b
420.3c
34.8
150.5
101152

75.9
20120
42.782.5
2766
45.9
227.0b
532.3c
53.7
195.9
151203
26.7

38.3996

C. beijerinckii BA101
C. beijerinckii P260
C. acetobutylicum JB200
Continuous
Fed-batch

C. acetobutylicum P262
C. acetobutylicum JB200
C. acetobutylicum P262
C. beijerinckii BA101

3.4/3.3
810/19.1
615/20.4
24.5/5.3
29.6/11.9
25.5/13.7

Reference

Ennis et al. (1986)

Maddox et al. (1995)
Ezeji et al. (2003)
Qureshi et al. (2008)
Mariano et al (2011)a
This study
Qureshi and Maddox (1991)
Xue et al. (2012)
Lu et al. (2012)
Qureshi et al. (1992)
Ezeji et al. (2004)
Ezeji et al. (2007b)

, Unavailable.
a
Vacuum was applied intermittently for 1.5 h at 4 h interval to remove ABE vapor.
b
Butanol and ABE in the condensate collected from the rst-stage gas stripping.
c
Butanol and ABE in the nal product collected from the two-stage gas stripping process.

in gas stripping in previous studies was relatively low, less than

120 g/L ABE or 70 g/L butanol (see Table 1), which still needs intensive energy for purication (Oudshoorn et al., 2009; Vane, 2008).
Moreover, additional gas such as nitrogen was also employed to
strip butanol, which increases the operation cost (Ezeji et al.,
2003; Ennis et al., 1986; Qureshi et al., 1992).
In conventional batch ABE fermentation, sugars in excess of
60 g/L could not be completely consumed due to strong butanol
inhibition (Ezeji et al., 2003). In the present study, an asporogenic
hyper-butanol producing C. acetobutylicum mutant JB200 (Lu
et al., 2012) was used to convert 80 g/L of glucose to ABE in
batch fermentation. With integrated two-stage gas stripping
using the fermentation off-gas (H2 and CO2) produced by the
microorganism, the butanol yield and productivity increased signicantly due to alleviated butanol inhibition. Furthermore, extremely high butanol and ABE concentrations of 420.3 and
532.3 g/L, respectively, were obtained in the nal product condensate, which were more than 20-fold higher than those obtained in conventional ABE fermentation. Thus, the total energy
consumption for butanol recovery from the fermentation integrated with the two-stage gas stripping process can be reduced
signicantly, providing an energy-efcient process for biobutanol
production.

2.2. Cell immobilization and bioreactor start-up

The spinner ask containing 1 L of the P2 medium was inoculated with 100 ml of actively growing cells (16 h) and then maintained at 37 C and pH 5.0, by the addition of 2 N NH4OH, and
agitated at 150 rpm for 2436 h until cell growth had reached an
optical density (OD6 0 0) of over 6.0. Then, the fermentation broth
was recirculated by a peristaltic pump through the brous bed bioreactor (FBB) for 2436 h for cells to be immobilized effectively
onto the brous matrix, as illustrated in Fig. 1. The FBB was made
of a glass column (50  400 mm, 250 mL working volume) packed
with spiral wound cotton towel and stainless steel wire cloth for a
total working volume of 1.0 L. After cell immobilization, the old
broth was drained out and fresh medium was fed to start batch
fermentation.
2.3. Batch fermentation
Batch fermentation was studied with the P2 medium in the FBB
system without gas stripping at 37 C and pH 5.0. Liquid samples
were withdrawn from the spinner ask periodically for the analysis of glucose, cell density and fermentation products.
2.4. Fermentation system coupled with two-stage gas stripping

2. Methods
2.1. Culture and media
Clostridium acetobutylicum JB200 derived from ATCC 55025 was
used in this study. The seed culture was prepared in the Clostridial
growth medium (CGM) containing 30 g/L glucose, 2 g/L yeast extract, 1 g/L Tryptone, minerals, and vitamins in a phosphate buffer
as described in Lu et al. (2012), and incubated at 37 C for 16 h
until active growth was observed. ABE fermentation was studied
using the P2 medium containing: glucose (80 g/L), yeast extract
(1 g/L), KH2PO4 (0.5 g/L), K2HPO4 (0.5 g/L), ammonium acetate
(2.2 g/L), vitamins (1 mg/L para-amino-benzoic acid, 1 mg/L thiamin and 0.01 mg/L biotin), and mineral salts (0.2 g/L MgSO4
7H2O,
0.01 g/L MnSO4
H2O, 0.01 g/L FeSO4
7H2O, 0.01 g/L NaCl), prepared
according to the procedures described previously (Qureshi and Blaschek, 1999). The media were sterilized by autoclaving at 121 C
and 15 psig for 30 min. All solutions were purged with nitrogen
for 1 h through a sterile 0.2 lm lter, either before or after
autoclaving.

Two condensers (Pyrex Graham coil, 30  300 mm, Fisher) for

vapor condensation, pumps and storage tanks were connected to
the FBB fermentation system illustrated in Fig. 1. The spinner ask,
FBB and condensers were autoclaved separately for 45 min, and
aseptically connected after sterilization. The whole system was
sparged with nitrogen before inoculation to ensure an oxygen-free
environment. For the rst-stage gas stripping, it was carried out by
circulating the off-gas at 1.5 L/min through the fermentation broth
and the condenser using a peristaltic pump, which was turned on
when butanol concentration in the fermentation broth increased to
8 g/L. The condenser was maintained at 2 C and the condensate
collected in a ask immersed in a cold water bath was pumped into
the storage tank for phase separation.
The organic phase consisting of 80% (v/v) butanol was transferred to the second-stage condensate storage tank, and the aqueous phase with 10% butanol was gas-stripped again. The secondstage gas stripping was conducted under the same conditions as
that for the rst-stage gas stripping. The condensate was collected
in the second-stage condensate storage tank and combined with

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C. Xue et al. / Bioresource Technology 135 (2013) 396402

Fig. 1. Experimental setup for ABE fermentation with two-stage gas stripping. 1, stirred tank fermentor; 2, brous bed bioreactor; 3, pH probe; 4, temperature probe; 5,
thermostat water inlet; 6, thermostat water outlet; 7, condenser; 8, rst-stage condensate storage tank; 9, cold-water bath; 10, second-stage condensate storage tank; 11,
cooling water inlet; 12, cooling water outlet; 13, peristaltic pump.

100

35
Glucose

Acetone

Butanol

Ethanol

ABE

Acetic

Butyric

OD

30

80

Biomass in the fermentation broth was estimated by measuring

the optical density at 600 nm with a spectrophotometer (UV-161,
Shimadzu, Columbia, MD). Glucose and products in the fermentation broth were assayed after removing cells by centrifugation at
13,200  g for 5 min. The glucose concentration was determined
with YSI 2700 Select Biochemistry Analyzer (Yellow Springs, Ohio).
Butanol, acetone, ethanol, acetic acid and butyric acid were determined using a gas chromatograph (GC-2014 Shimadzu, Columbia,
MD) equipped with a ame ionization detector (FID) and a fused
silica column (Stabilwax-DA, 30 m long, 0.25 lm lm thickness
and 0.25 mm ID, Restek, Bellefonte, PA) following the method previously described (Yu et al., 2011).

25

Glucose, g/L

2.5. Analytical methods

15

40

10
5
0

10

20

30

40

50

60

Time, h
35

B 100

Glucose
ABE

Acetone
Acetic

Butanol
Butyric

Ethanol
OD

30

80
25

Gas stripping

Glucose, g/L

Batch ABE fermentations without and with in situ gas stripping

for product recovery were studied with P2 medium containing
80.0 g/L glucose, and the results are shown in Fig. 2. Without
gas stripping, 7.9 g/L acetone, 16.2 g/L butanol and 1.4 g/L ethanol
were produced from 80 g/L glucose in 53 h, resulting in a total
ABE of 25.5 g/L. The nal concentrations of acetic, butyric and total
acids were 1.2, 0.9, and 2.1 g/L, respectively. In contrast, with in situ
gas stripping, which was initiated at 28 h, the fermentation time
was shortened to 48 h and more ABE (31.8 g/L; 10.3 g/L acetone,
19.8 g/L butanol and 1.7 g/L ethanol) were produced from the same
amount of glucose (80 g/L). The nal concentrations of acetic, butyric, and total acids were 0.4, 1.7, and 2.1 g/L, respectively. The increased butanol and ABE production resulted in increased
butanol and ABE yields, from 0.20 to 0.25 g/g and 0.32 to 0.40 g/
g, respectively, and butanol and ABE productivities, from 0.30 g/
L h to 0.41 g/L h and 0.48 g/L h to 0.66 g/L h, respectively. These results are summarized in Table 2. The improved ABE fermentation
with in situ gas stripping can be attributed to the removal of butanol from the fermentation broth, alleviating butanol toxicity and
inhibition to cells.

20

20

3. Results and discussion

3.1. Enhanced butanol fermentation by in situ gas stripping

60

Products, g/L; OD 600

60

20
15

40

10
20

Products, g/L; OD 600

the organic phase from the rst-stage stripping for further product
purication. Liquid samples were taken from the spinner ask, and
the rst- and second-stage condensate storage tanks for analysis.

5
0

10

20

30

40

50

Time, h
Fig. 2. Time course proles of ABE fermentation by C. acetobutylicum JB200
immobilized in a brous bed bioreactor. (A) Batch fermentation without in situ gas
stripping; (B) Batch fermentation with in situ gas stripping starting at 28 h. Dashed
lines indicate the total production including products collected in the gas stripping
condensate and those remained in the fermentation broth.

It is noted that a hyper-butanol producing C. beijerinckii BA101

could produce up to 20.9 g/L butanol, which was the highest butanol titer ever reported for ABE fermentation and represented a

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C. Xue et al. / Bioresource Technology 135 (2013) 396402

Table 2
Comparison of batch ABE fermentations of glucose by C. acetobutylicum JB200 without
and with in situ gas stripping.
Fermentation
parameters

Batch fermentation
without gas stripping

Batch fermentation
with gas stripping

Initial glucose, g/L

Residual glucose, g/L
Maximum OD
Fermentation time, h
Acetone, g/L
Butanol, g/L
Ethanol, g/L
Total ABE, g/L
Butanol yield, g/g
ABE yield, g/g
Butanol productivity, g/L
h
ABE productivity, g/L
h
Acetic acid, g/L
Butyric acid, g/L
Total acids, g/L

80.0 1.1
0.3 0.1
1.30
53
7.9 0.1
16.2 0.9
1.4 0.2
25.5 1.2
0.20 0.01
0.32 0.02
0.30 0.02
0.48 0.02
1.2 0.1
0.9 0.1
2.1 0.2

80.0 1.1
0
1.64
48
10.3 0.2
19.8 0.8
1.7 0.1
31.8 1.1
0.25 0.01
0.40 0.01
0.41 0.02
0.66 0.02
0.4 0.1
1.7 0.1
2.1 0.2

74% increase from 12 g/L usually obtained in ABE fermentation

(Chen and Blaschek, 1999; Ezeji et al., 2004; Ezeji et al., 2007b).
However, the strains capability for butanol production was still severely inhibited by butanol toxicity, which could only be alleviated
by in situ removal of butanol. As demonstrated in this and previous
studies (Ezeji et al., 2003; Ezeji et al., 2007b; Lu et al., 2012; Maddox et al., 1995; Qureshi et al., 2008), gas stripping is highly efcient for in situ butanol recovery and can effectively alleviate
butanol inhibition and increase ABE fermentation productivity
(see Table 1). However, the butanol concentration obtained in
gas stripping in previous studies was almost always less than
80 g/L or the solubility of butanol in water, which does not allow
phase separation or liquid demixing to occur. Consequently, gas
stripping has not been considered as an energy-saving butanol
recovery method until recently (Lu et al., 2012; Xue et al., 2012),
and its potential in energy saving for biobutanol production has
not been fully explored nor optimized. In our recent studies, we
demonstrated that under proper conditions gas stripping can produce condensate with highly concentrated butanol (Lu et al., 2012;
Xue et al., 2012). In this study, we further studied a two-stage gasstripping process to further explore its energy-saving potential for
butanol recovery.
3.2. Two-stage gas stripping for butanol recovery from fermentation
broth
The rst-stage gas stripping was used to remove ABE from the
fermentation broth to alleviate their inhibition on cells as well as
enhance butanol production, which has been discussed above.
The second-stage gas stripping was applied to further concentrate

butanol from the aqueous phase formed with the condensate collected from the rst-stage gas stripping. The compositions of the
condensates collected from the rst and second-stage gas stripping
are summarized in Table 3.
The solubility of butanol in water at 20 C is 7.7% (w/w) and
phase separation occurs spontaneously when butanol concentration is higher than 8%. The condensate of the rst-stage gas stripping contained 42.7 g/L acetone, 175.6 g/L butanol, 8.6 g/L
ethanol, 0.4 g/L acetic acid, and 0.8 g/L butyric acid. The butanol
concentration in the condensate was high enough for effective
phase separation, generating an organic phase containing 39.3 g/L
acetone, 612.3 g/L butanol and 9.1 g/L ethanol, and an aqueous
phase containing 43.4 g/L acetone, 101.3 g/L butanol, 8.5 g/L ethanol, 0.5 g/L acetic acid, and 0.7 g/L butyric acid. It is noted that no
acid was present in the organic phase and the butanol concentration in the aqueous phase was higher than the butanol solubility
in water because of the presence of acetone. The second gas stripping of the aqueous phase produced a highly concentrated condensate with 118.7 g/L acetone, 336.6 g/L butanol, and 22.1 g/L
ethanol, which after combining with the organic phase from the
rst-stage gas stripping gave a nal product mixture of 94.0 g/L
acetone, 420.3 g/L butanol, 18.0 g/L ethanol, and no acid. When
butanol is concentrated to such a high level, energy consumption
in nal product recovery as fuel or solvent by existing technologies
such as distillation can be reduced signicantly. Gas stripping is
not an energy-intensive process, as little energy is required in
pumping the stripping gas through the fermentor and the cooling
water through the condenser for recovering the solvent vapors.
Compared to steam stripping and distillation used in the conventional ABE fermentation recovery process, the two-stage gas stripping process would consume much less energy because only a
small amount of water needs to be removed in the nal dewatering
and product purication step.

3.3. Dynamics of the second-stage gas stripping

While the rst-stage gas stripping was controlled by butanol
concentration within the fermentation system and thus the operation was relatively stable, the second-stage gas stripping of the
aqueous phase collected from the condensate of the rst-stage
gas stripping was operated in a batch mode at unsteady state. Thus,
the dynamics of the second-stage gas stripping was further studied
using a simulated ABE solution because the volume of the aqueous
phase collected from the rst-stage gas stripping was not enough
to perform the study. As can be seen in Fig. 3, butanol and acetone
concentrations decreased drastically as gas stripping was ongoing,
but the ethanol concentration only dropped slightly, indicating an
excellent selectivity for butanol and acetone by the gas stripping,
particularly for butanol. Meanwhile, butanol and acetone

Table 3
Product compositions and volumes obtained from rst and second stage gas stripping.
First-stage gas stripping

Acetone, g/L
Butanol, g/L
Ethanol, g/L
ABE, g/L
Acetic acid, g/L
Butyric acid, g/L
Total acids, g/L
Volume, mL
a
b
+

Condensate

Organic phase

Aqueous phase

42.7 3.2
175.6 12.9
8.6 0.9
226.9 17.0
0.4 0.02
0.8 0.03
1.2 0.05
350 1

39.3 4.8
612.3 40.8
9.1 1.2
660.7 46.8
None
None
None
50 1

43.4 2.1
101.3 5.4+
8.5 0.4
153.2 7.9
0.5 0.03
0.7 0.03
1.2 0.06
300 1

Condensate from second-stage gas strippinga

Final product mixtureb

118.7 4.3
336.6 17.4
22.1 2.3
477.4 24.0
None
None
None
75 1

94.0 5.6
420.3 23.5
18.0 2.1
532.3 31.2
None
None
None
125 1

Using the simulated feed solution formulated based on the composition of the aqueous phase obtained in the rst-stage gas stripping.
The mixture of the organic phase from the rst-stage gas stripping and the condensate from the second-stage gas stripping.
Butanol concentration was higher than its solubility (77 g/L) in water due to the presence of acetone.

C. Xue et al. / Bioresource Technology 135 (2013) 396402

A 120

500

80

400

Butanol in solution
Acetone in solution
Ehanol in solution
Butanol in condensate
Acetone in condensate

60
40

300
200

Ethanol in condensate

20

ABE in condensate, g/L

ABE in solution, g/L

100

600

12

15

18

Ethanol
12

Time, h

20

40

60

80

100

ABE in solution, g/L

B 600

30

500

25
400

Selectivity

ABE in condensate, g/L

Acetone

16

100

20
Butanol

Stripping rate, g/L/h

400

300
Butanol

200

Acetone
Ethanol

100

20
15
10
Butanol

20

40

60

80

100

Acetone

Ethanol

ABE in solution, g/L

0
Fig. 3. Kinetics of gas stripping of ABE solution. (A) Time-courses of gas stripping of
ABE solution showing changes in ABE concentrations in the feed solution and in the
condensate. (B) ABE concentrations in the condensate from gas striping as affected
by ABE concentrations in the feed solution. The feed solution has an initial ABE
composition that was the same as that from the aqueous phase collected from the
condensate of the in situ gas stripping of the batch fermentation shown in Fig. 2B.

concentrations in the condensate also decreased gradually with

time because of the reduced butanol and acetone concentrations
in the feed solution. It should be noted that the nal condensate
obtained in this experiment contained 336.6 g/L of butanol,
118.7 g/L of acetone, and 22.1 g/L of ethanol (Table 3), which were
lower than the calculated data (386.4 g/L of butanol, 134.5 g/L of
acetone, and 23.2 g/L of ethanol) shown in Fig. 3, which were based
on the material balance using the ABE concentrations in the feed
solution and volume changes over time during the gas stripping
process. Based on the initial ABE concentrations (101.3 g/L of butanol, 43.4 g/L of acetone and 8.5 g/L of ethanol) and nal ABE concentrations (3.7 g/L of butanol, 7.7 g/L of acetone and 3.9 g/L of
ethanol) in the feed solution, the recovery rate of butanol, acetone
and ethanol for the gas stripping were 85.4%, 78.9% and 99.1%,
respectively. The differences or less than 100% recovery can be
mainly attributed to loss in liquid sampling and adsorption to tubing and condenser in the stripping system.
The performance and efciency of gas stripping are greatly affected by the ABE concentrations in the feed solution. As can be
seen in Fig. 3B, there is a strong correlation between the ABE concentrations in the condensate and in the feed solution. When butanol concentration in the feed solution is below 5 g/L, gas stripping
is no longer efcient and butanol concentration in the condensate
would decrease signicantly due to a larger amount of water being
stripped with butanol. Furthermore, as shown in Fig. 4, the gas
stripping rate increased with increasing the feed concentration.

20

40

60

80

100

ABE in solution, g/L

Fig. 4. Effects of ABE concentrations in the feed solution on gas stripping rate and
selectivity. (A) Stripping rate; (B) selectivity. Dashed lines indicate the average ABE
selectivity values in the ABE concentration ranges studied.

In addition, the selectivity for butanol, which is dened as the concentration ratio between the condensate and feed solution, is much
higher than those for acetone and ethanol, and is generally higher
than 10. Clearly, gas stripping is efcient for butanol recovery
when the butanol concentration is higher than 510 g/L, allowing
phase separation in the condensate to result in a highly concentrated butanol solution.
However, the butanol concentration in the condensate should
be as high as possible for the energy efciency of further butanol
purication and dewatering by distillation or other technologies.
Therefore, in order to capture the full energy-saving potential of
gas stripping, a second-stage gas stripping of the aqueous phase
from the rst-stage gas stripping is necessary. As demonstrated
in the present study, second-stage gas stripping can recover more
than 80% of the butanol (100 g/L) present in the rst-stage
aqueous phase and produce a condensate with more than 330 g/L
of butanol. The more than threefold increase in the butanol concentration would reduce at least 67% of the energy required in
the nal dewatering of biobutanol for fuel application. Overall,
gas stripping can increase butanol concentration from less than
2% (w/v) to over 40% (w/v) and the energy saving thus can be more
than 90% compared to conventional ABE fermentation without gas
stripping. It is thus desirable to use two-stage gas stripping for
butanol recovery to further reduce energy consumed in biobutanol
production.

C. Xue et al. / Bioresource Technology 135 (2013) 396402

401

3.4. Comparison to other ABE fermentation studies with gas stripping

References

Butanol fermentation coupled with gas stripping has been

widely studied, and a literature survey is summarized in Table 1.
As can be seen in Table 1, due to butanol toxicity, almost all ABE
fermentations with gas stripping, except for our own recent studies, were conducted at a low butanol concentration of 28 g/L, and
the condensates contained less than 70 g/L butanol, not enough for
phase separation, making further butanol purication still energyintensive as it would require about 1431 MJ/kg butanol produced (Xue et al., 2012). In this study, gas stripping was operated
at much higher butanol concentrations of 814 g/L, producing the
condensate with 175.6 g/L butanol or 227.0 g/L ABE for efcient
phase separation. The more than twofold increase in the butanol
titer (175.6 g/L vs. <70 g/L) in the condensate should reduce at
least 50% of the energy (715 MJ/kg) required for further purication. Furthermore, the ABE present in the aqueous phase after
phase separation can be further concentrated in the second gas
stripping to more than 336 g/L butanol. After combining with
the upper phase of the rst stripping condensate, the overall butanol concentration would be 420 g/L, which is the highest ever reported for any process for biobutanol production. This could
further reduce energy consumption in butanol purication to less
than 5 MJ/kg butanol. It should be noted that conventional distillation requires 36 MJ/kg butanol from a solution containing 1%
(w/v) butanol (Matsumura et al., 1988), which would have no
net energy gain as the energy content of butanol is 36 MJ/kg. To
our best knowledge, this is the rst study demonstrating the
operation of two-stage gas stripping at high butanol concentration
during ABE fermentation with a purpose of harvesting condensate
with extremely high butanol concentration to reduce energy
consumption by more than 85% for butanol purication as a fuel
or solvent. Although other butanol recovery techniques, including
pervaporation, extraction and adsorption, have also been evaluated for their potential to reduce energy consumption in ABE
fermentation (Oudshoorn et al., 2009), none of them have shown
similar performance to the two-stage gas stripping demonstrated
in the present study.

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4. Conclusions
A two-stage gas stripping process was developed for in situ
butanol recovery from ABE fermentation. The rst-stage gas stripping was initiated at butanol concentration above 8 g/L, which not
only alleviated butanol inhibition, but also made the condensate
containing enough butanol for efcient phase separation. After
the organic phase was collected, the aqueous phase was subjected
to second-stage gas stripping, resulting in a nal product mixture
containing more than 400 g/L butanol (500 g/L ABE), which was
20-fold higher than that achieved in conventional ABE fermentation. Therefore, this process can signicantly reduce energy consumption in nal product recovery.

Acknowledgements
This work was supported in part by the Ohio Department of
Development-Third Frontier Advanced Energy Program (Tech 08036), the National Science Foundation STTR program (IIP0810568, IIP-1026648), and Advanced Research Projects AgencyEnergy (DE-AR0000095). Financial support from the Fundamental
Research Funds for the Central Universities (DUT11RC(3)77), China
Postdoctoral Science Foundation (20110491527) and China Scholarship Council (2009100607) is also acknowledged.

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