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Frontiers of Chemical Science and Engineering

ISSN 2095-0179

ISSN 2095-0187(Online)

CN 11-5981/TQ

邮发代号 80-969

2019 Impact Factor: 3.552

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Frontiers of Chemical Science and Engineering  2015, Vol. 9 Issue (3): 369-375   https://doi.org/10.1007/s11705-015-1525-3
  本期目录
High production of butyric acid by Clostridium tyrobutyricum mutant? ?
Chao Ma,Jianfa Ou,Matthew Miller,Sarah McFann,Xiaoguang (Margaret) Liu()
Department of Chemical and Biological Engineering, The University of Alabama, 245 7th Avenue, Tuscaloosa, AL 35401, USA
 全文: PDF(713 KB)   HTML
Abstract：

The objective of this study was to improve the production of butyric acid by process optimization using the metabolically engineered mutant of Clostridium tyrobutyricum (PAK-Em). First, the free-cell fermentation at pH 6.0 produced butyric acid with concentration of 38.44 g/L and yield of 0.42 g/g. Second, the immobilized-cell fermentations using fibrous-bed bioreactor (FBB) were run at pHs of 5.0, 5.5, 6.0, 6.5 and 7.0 to optimize fermentation process and improve the butyric acid production. It was found that the highest titer of butyric acid, 63.02 g/L, was achieved at pH 6.5. Finally, the metabolic flux balance analysis was performed to investigate the carbon rebalance in C. tyrobutyricum. The results show both gene manipulation and fermentation pH change redistribute carbon between biomass, acetic acid and butyric acid. This study demonstrated that high butyric acid production could be obtained by integrating metabolic engineering and fermentation process optimization.
Key words： Clostridium tyrobutyricum    butyric acid production    fermentation    mutant    pH    flux balance analysis
收稿日期: 2015-05-02      出版日期: 2015-09-30
Corresponding Author(s): Xiaoguang (Margaret) Liu   
 引用本文:   
. [J]. Frontiers of Chemical Science and Engineering, 2015, 9(3): 369-375.
Chao Ma,Jianfa Ou,Matthew Miller,Sarah McFann,Xiaoguang (Margaret) Liu. High production of butyric acid by Clostridium tyrobutyricum mutant? ?. Front. Chem. Sci. Eng., 2015, 9(3): 369-375.
 链接本文:  
https://academic.hep.com.cn/fcse/CN/10.1007/s11705-015-1525-3
https://academic.hep.com.cn/fcse/CN/Y2015/V9/I3/369
Strains Fermentation mode Sugar Concentration /g·L−1 Ref.
C. populeti Batch, free-cell Glucose 6.30 [11]
C. butyricum ZJUCB Batch, free-cell Glucose 12.25 [12]
Fed-batch, free-cell 16.74
C. butyricum S21 Batch, free-cell Lactose 18.60 [13]
C. beijerinckii Batch, free-cell Lactose 12.00 [14]
C. thermobutyricum Fed-batch, free-cell Glucose 19.40 [15]
C. tyrobutyricum JM1 Batch, free-cell Glucose 13.76 [16]
C. tyrobutyricum, wild type Fed-batch, free-cell Glucose 24.88 [17]
Continuous, free-cell Glucose 33.00 [18]
C. tyrobutyricum, mutant Fed-batch, immobilized-cell Glucose 49.90 [19]
Repeated fed-batch, immobilized-cell Glucose 86.9 [20]
E. coli Batch, free-cell Glucose 10.00 [21]
Tab.1  
Reaction No. Biological function Stoichiometric equation a,b)
(1) Biomass formation 2 Glucose+ 1.75 NADH+ 1.75 H+ + 29.7 ATP →3 C4H6.4O1.72N+ 1.75 NAD+ + 29.7 ADP+ 29.7 Pi
(2) Formation of pyruvate (glycolysis) Glucose+ 2 NAD+ + 2 ADP+ 2 Pi → 2 Pyruvate+ 2 NADH+ 2 H+ + 2 ATP
(3) Formation of AcCoAand CO2 Pyruvate+ CoA+ Fdox → AcCoA+ Fdred + CO2
(4) Formation of H2 Fdred+ 2 H+ → H2+ Fdox
(5) Formation of NADH Fdred+ NAD+ ⇔ NADH+ H+ + Fdox
(6) Formation of acetate AcCoA+ ADP+ Pi ⇔ Acetate+ CoA+ ATP
(7) Formation of BuCoAand water 2 AcCoA+ 2 NADH+ 2H+ → BuCoA+ 2 NAD+ + CoA+ H2O
(8) Formation of butyrateor acetate BuCoA+ Acetate ⇔ Butyrate+ AcCoA
(9) Formation of butyrate BuCoA+ ADP+ Pi ⇔ Butyrate+ CoA+ ATP
Tab.2  
Fig.1  
Products Wild type (control) PAK-Em
Cell growth Growth rate µ /h−1 0.21±0.01 0.14±0.01
Biomass yield /(g·g−1) 0.06±0.01 0.04±0.01
Butyric acid Concentration /(g·L−1) 19.24±0.05 38.44±0.03
Yield /(g·g−1) 0.34±0.02 0.42±0.01
Acetic acid Concentration /(g·L−1) 4.22±0.002 7.16±0.002
Yield /(g·g−1) 0.07±0.001 0.07±0.01
C4/C2 B/A ratio /(g·g−1) 4.56±0.85 5.36±0.61
Tab.3  
Fig.2  
Products pH
5.0 5.5 6.0 6.5 7.0
Butyrate Conc. /(g·L−1) 14.79±0.99 23.18±0.78 50.11±2.42 63.02±1.54 61.01±0.78
Yield /(g·g−1) 0.37±0.03 0.38±0.01 0.45±0.02 0.45±0.01 0.42±0.01
Acetate Conc. /(g·L−1) 2.11±0.02 3.13±0.02 7.03±0.01 7.26±0.03 7.09±0.02
Yield /(g·g−1) 0.03±0.004 0.03±0.006 0.08±0.01 0.05±0.01 0.04±0.01
C4/C2 Ratio /(g·g−1) 6.53 6.77 7.12 8.60 8.60
Tab.4  
Fig.3  
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