Disruption of the acetoacetate decarboxylase gene in solvent-producing Clostridium acetobutylicum increases the butanol ratio

被引:176
作者
Jiang, Yu [1 ,2 ]
Xu, Chongmao [1 ,2 ]
Dong, Feng [1 ,2 ]
Yang, Yunliu [1 ]
Jiang, Weihong [1 ,2 ]
Yang, Sheng [1 ,2 ,3 ]
机构
[1] Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Plant Physiol & Ecol, Key Lab Synthet Biol, Shanghai 200032, Peoples R China
[2] Shanghai Res & Dev Ctr Ind Biotechnol, Shanghai 201201, Peoples R China
[3] Chinese Acad Sci, Huzhou Ctr Ind Biotechnol, Shanghai Inst Biol Sci, Huzhou 313000, Peoples R China
基金
中国国家自然科学基金;
关键词
Acetone; Acetoacetic acid decarboxylase; Butanol ratio; Clostridium acetobutylicum; Gene disruption; ENHANCED BUTYRIC-ACID; ESCHERICHIA-COLI; DOWN-REGULATION; ATCC; 824; ACETONE FORMATION; ETHANOL FERMENTATION; HYDROGEN-PRODUCTION; NADH AVAILABILITY; COA-TRANSFERASE; CARBON-MONOXIDE;
D O I
10.1016/j.ymben.2009.06.002
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
A possible way to improve the economic efficacy of acetone-butanol-ethanol fermentation is to increase the butanol ratio by eliminating the production of other by-products, such as acetone. The acetoacetate decarboxylase gene (adc) in the hyperbutanol-producing industrial strain Clostridium acetobutylicum EA 2018 was disrupted using TargeTron technology. The butanol ratio increased from 70% to 80.05%, with acetone production reduced to approximately 0.21 g/L in the adc-disrupted mutant (2018adc). pH control was a critical factor in the improvement of cell growth and solvent production in strain 2018adc. The regulation of electron flow by the addition of methyl viologen altered the carbon flux from acetic acid production to butanol production in strain 2018adc, which resulted in an increased butanol ratio of 82% and a corresponding improvement in the overall yield of butanol from 57% to 70.8%. This study presents a general method of blocking acetone production by Clostridium and demonstrates the industrial potential of strain 2018adc. (C) 2009 Elsevier Inc. All rights reserved.
引用
收藏
页码:284 / 291
页数:8
相关论文
共 48 条
[31]   Engineering solventogenic clostridia [J].
Papoutsakis, Eleftherios T. .
CURRENT OPINION IN BIOTECHNOLOGY, 2008, 19 (05) :420-429
[32]   A comparative genomic view of clostridial sporulation and physiology [J].
Paredes, CJ ;
Alsaker, KV ;
Papoutsakis, ET .
NATURE REVIEWS MICROBIOLOGY, 2005, 3 (12) :969-978
[33]   Butanol production from wheat straw hydrolysate using Clostridium beijerinckii [J].
Qureshi, Nasib ;
Saha, Badal C. ;
Cotta, Michael A. .
BIOPROCESS AND BIOSYSTEMS ENGINEERING, 2007, 30 (06) :419-427
[34]   ALTERED ELECTRON FLOW IN CONTINUOUS CULTURES OF CLOSTRIDIUM-ACETOBUTYLICUM INDUCED BY VIOLOGEN DYES [J].
RAO, G ;
MUTHARASAN, R .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1987, 53 (06) :1232-1235
[35]   Effect of different levels of NADH availability on metabolic fluxes of Escherichia coli chemostat cultures in defined medium [J].
Sáanchez, AM ;
Bennett, GN ;
San, KY .
JOURNAL OF BIOTECHNOLOGY, 2005, 117 (04) :395-405
[36]   Targeted gene disruption by use of a group II intron (targetron) vector in Clostridium acetobutylicum [J].
Shao, Lijun ;
Hu, Shiyuan ;
Yang, Yi ;
Gu, Yang ;
Chen, Jun ;
Yang, Yunliu ;
Jiang, Weihong ;
Yang, Sheng .
CELL RESEARCH, 2007, 17 (11) :963-965
[37]   Metabolic engineering of Escherichia coli for 1-butanol and 1-propanol production via the keto-acid pathways [J].
Shen, C. R. ;
Liao, J. C. .
METABOLIC ENGINEERING, 2008, 10 (06) :312-320
[38]   Metabolic engineering of the non-sporulating, non-solventogenic Clostridium acetobutylicum strain M5 to produce butanol without acetone demonstrate the robustness of the acid-formation pathways and the importance of the electron balance [J].
Sillers, Ryan ;
Chow, Alison ;
Tracy, Bryan ;
Papoutsakis, Eleftherios T. .
METABOLIC ENGINEERING, 2008, 10 (06) :321-332
[39]   Aldehyde-Alcohol Dehydrogenase and/or Thiolase Overexpression Coupled With CoA Transferase Downregulation Lead to Higher Alcohol Titers and Selectivity in Clostridium acetobutylicum Fermentations [J].
Sillers, Ryan ;
Al-Hinai, Mohab Ali ;
Papoutsakis, Eleftherios T. .
BIOTECHNOLOGY AND BIOENGINEERING, 2009, 102 (01) :38-49
[40]  
Soucaille P, 2008, Process for the biological production of n-butanol at high yield, Patent No. [WO/2008/052973, 2008052973]