Impact of deletion of the genes encoding acetate kinase on production of L-tryptophan by Escherichia coli

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• 作者：Chunguang Zhao ; LiKun Cheng ; Jian Wang ; Zhiqiang Shen ; Ning Chen
• 关键词：Escherichia coli ; L ; Tryptophan ; Acetate kinase ; ackA ; tdcD ; Metabolic flux
• 刊名：Annals of Microbiology
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：66
• 期：1
• 页码：261-269
• 全文大小：888 KB
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• 作者单位：Chunguang Zhao (1)
  LiKun Cheng (2)
  Jian Wang (3)
  Zhiqiang Shen (2)
  Ning Chen (1)
  
  1. College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, People’s Republic of China
  2. Shandong Binzhou Animal Propolis Vaccine Research & Development Center, Shandong Lvdu Bio-science & Technology Co.Ltd., Shandong Binzhou Animal Science & Veterinary Medicine Academy, Binzhou, 256600, Shandong, People’s Republic of China
  3. College of Biological and Agricultural Engineering, Jilin University, Changchun, 130022, People’s Republic of China
  
• 刊物主题：Microbiology; Microbial Genetics and Genomics; Microbial Ecology; Fungus Genetics; Medical Microbiology; Applied Microbiology;
• 出版者：Springer Berlin Heidelberg
• ISSN：1869-2044

文摘

The production of L-tryptophan was increased by reducing acetate accumulation through a decrease in acetate kinase activity by gene deletion. The effects of disruption of the genes for acetate kinase (ackA) and an enzyme with propionate/acetate kinase activity (tdcD) on L-tryptophan production were investigated. The ackA and/or tdcD deletion mutants accumulated less acetate and more L-tryptophan than the parental strain. Furthermore, the production of L-tryptophan obtained with ackA-tdcD mutant were more than the mutants with a single deletion of ackA or tdcD, while higher production of L-tryptophan and lower concentration of acetate were accumulated in the ackA mutant than the mutant with a lesion in tdcD. In L-tryptophan fed-batch fermentation using the ackA-tdcD mutant, the excretion of acetate was reduced to 1.22 g/L, a 21.79 % reduction compared with the parental strain, and the production of L-tryptophan and glucose conversion rate were increased to 52.5 g/L and 47.9 g/L, respectively, which represented 6.49 % and 10.88 % increases compared with the parental strain, and the glucose conversion rate reached a high level of 21.2 %, which was 8.16 % higher than the parental strain. In addition, the metabolic flux analysis of TRTH and TRTHAT indicated that the carbon flux through EMP was decreased by 8.37 % and the carbon flux through PP was increased by 57.03 % in TRTHAT compared with TRTH. The flux of acetate and tryptophan formation of TRTHAT were 5.2 % and 17.3 %, which were 3.67-times lower and 1.75-times higher than these of TRTH, respectively. Keywords Escherichia coli L-Tryptophan Acetate kinase ackA tdcD Metabolic flux