Pull-in urea cycle for the production of fumaric acid in Escherichia coli

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• 作者：Ting Zhang ; Zening Wang ; Li Deng ; Tianwei Tan…
• 关键词：Fumaric acid ; Escherichia coli ; Urea cycle ; Metabolic analysis
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2015
• 出版时间：June 2015
• 年：2015
• 卷：99
• 期：12
• 页码：5033-5044
• 全文大小：1,296 KB
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• 作者单位：Ting Zhang (1)
  Zening Wang (1)
  Li Deng (1) (2)
  Tianwei Tan (1)
  Fang Wang (1)
  Yajun Yan (3)
  
  1. Beijing Bioprocess Key Laboratory, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, People’s Republic of China
  2. Amoy—BUCT Industrial of Bio-technovation Institute, Amoy, 361022, People’s Republic of China
  3. Biochemical Engineering Program, College of Engineering, University of Georgia, Athens, GA, 30602, USA
  
• 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

Fumaric acid (FA) is an important raw material in the chemical and pharmaceutical industries. In this work, Escherichia coli was metabolically engineered for the production of FA. The fumA, fumB, fumC, and frdABCD genes were deleted to cut off the downstream pathway of FA. In addition, the iclR and arcA genes were also deleted to activate the glyoxylate shunt and to reinforce the oxidative Krebs cycle. To increase the FA yield, this base strain was further engineered to be pulled in the urea cycle by overexpressing the native carAB, argI, and heterologous rocF genes. The metabolites and the proteins of the Krebs cycle and the urea cycle were analyzed to confirm that the induced urea cycle improved the FA accumulation. With the induced urea cycle, the resulting strain ABCDIA-RAC was able to produce 11.38?mmol/L of FA from 83.33?mmol/L of glucose in a flask culture during 24?h of incubation.