Biosynthesis of butenoic acid through fatty acid biosynthesis pathway in Escherichia coli

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• 作者：Xiping Liu ; Haiying Yu ; Xu Jiang ; Guomin Ai…
• 关键词：Butenoic acid ; Escherichia coli ; Bacteroides thetaiotaomicron thioesterase ; Fatty acid biosynthesis
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2015
• 出版时间：February 2015
• 年：2015
• 卷：99
• 期：4
• 页码：1795-1804
• 全文大小：683 KB
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• 作者单位：Xiping Liu (1) (3)
  Haiying Yu (1)
  Xu Jiang (1)
  Guomin Ai (2)
  Bo Yu (1)
  Kun Zhu (1)
  
  1. CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China
  3. University of Chinese Academy of Sciences, Beijing, 100049, China
  2. State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0614

文摘

Butenoic acid is a C4 short-chain unsaturated fatty acid mainly used in the preparation of resins, pharmaceuticals, and fine chemicals. However, butenoic acid derived from petroleum is costly and unfriendly to the environment. Here, we report a novel biosynthetic strategy to produce butenoic acid by utilizing the intermediate of fatty acid biosynthesis pathway in engineered Escherichia coli. A thioesterase gene (B. thetaiotaomicron thioesterase (bTE)) from Bacteroides thetaiotaomicron was heterologously expressed in E. coli to specifically convert butenoyl-acyl carrier protein (ACP), a fatty acid biosynthesis intermediate, to butenoic acid. The titer of butenoic acid ranged from 0.07 to 11.4?mg/L in four different E. coli strains with varied expressing vectors. Deletion of endogenous fadD gene (encoding acyl-CoA synthetase) to block fatty acid oxidation improved the butenoic acid production in all strains to some extent. The highest butenoic acid accumulation of 18.7?mg/L was obtained in strain XP-2 (BL21-?em class="a-plus-plus">fadD/pET28a-bTE). Moreover, partially inhibiting the enoyl-ACP reductase (FabI) of strain XP-2 by triclosan increased butenoic acid production by threefold, and the butenoic acid titer was further increased to 161.4?mg/L by supplying glucose and tryptone in the M9 medium. Fed-batch fermentation of this strain further enhanced butenoic acid production to 4.0?g/L within 48?h. The butenoic acid tolerance assay revealed that this strain could tolerate 15-0?g/L of butenoic acid.