Food and Fermentation Industries >

2023 , Vol. 49 >Issue 21: 9 - 15

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.034924

Systems metabolic engineering of Corynebacterium glutamicum for the efficient production of L-alanine

  • WU Chen ,
  • WANG Zeting ,
  • ZHAO Guihong ,
  • LYU Gengcheng ,
  • WANG Feiao ,
  • LIU Yuexiang ,
  • CHEN Ning ,
  • LI Yanjun
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  • 1(College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China)
    2(Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin University of Science and Technology, Tianjin 300457, China)
    3(National and Local United Engineering Lab of Metabolic Control Fermentation Technology, Tianjin University of Science and Technology, Tianjin 300457, China)

Received date: 2023-01-17

  Revised date: 2023-02-16

  Online published: 2023-12-08

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Abstract

L-alanine is an important natural amino acid, which is widely used in medicine, food and feed industries. At present, L-alanine is mainly produced by Escherichia coli. It is of great significance to develop safe and industrially robust Corynebacterium glutamicum production strains. In this study, exogenous alanine dehydrogenase was optimized to synthesize L-alanine efficiently without using other amino acids as an amino donor. The non-phosphotransferase system glucose uptake pathway was strengthened by the simultaneous inactivation of the transcriptional repressor IolR and overexpression of the glucose kinase, which drastically promote the cell growth and L-alanine synthesis. The introduction of exogenous Entner-Doudoroff pathway that supply pyruvate only through 4-step reactions, an alternative to the 10-step Embden-Meyerhof-Parnas (EMP) pathway, further increased the production of L-alanine. In addition, in order to avoid the use of antibiotics and guarantee the genetic stability of the producer, essential genes were integrated into the genome of the strain. The final engineered strain produced 104 g/L L-alanine after 48 h fermentation in a 5 L fermenter. The study lays a solid foundation for the industrial production of L-alanine by C. glutamicum, and provides a reference for the metabolic engineering of other microbes to produce products derived from pyruvate.

Key words： Corynebacterium glutamicum; L-alanine; alanine dehydrogenase; non-phosphotransferase system; embden-meyerhof-parnas (EMP) pathway; entner-doudoroff pathway

Cite this article

WU Chen , WANG Zeting , ZHAO Guihong , LYU Gengcheng , WANG Feiao , LIU Yuexiang , CHEN Ning , LI Yanjun . Systems metabolic engineering of Corynebacterium glutamicum for the efficient production of L-alanine[J]. Food and Fermentation Industries, 2023 , 49(21) : 9 -15 . DOI: 10.13995/j.cnki.11-1802/ts.034924

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