Food and Fermentation Industries >

2026 , Vol. 52 >Issue 14: 48 - 56

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.044858

Systematic metabolic engineering of Escherichia coli for high-yield L-tryptophan production

  • WU Yadong ,
  • YANG Yan ,
  • WANG Jing ,
  • ZHANG Weifeng ,
  • WANG Xiaonan ,
  • CHEN Ning
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  • 1(College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China)
    2(Ruzhou Kunhe Biotechnology Co.Ltd., Ruzhou 467500, China)

Received date: 2025-10-17

  Revised date: 2025-12-17

  Online published: 2026-08-21

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Abstract

L-Tryptophan (L-Trp) is one of the essential amino acids for humans and animals, widely used in food additives, animal feed, and pharmaceutical applications.Due to its advantages of high efficiency and low cost, microbial fermentation has become the mainstream method for L-tryptophan production.Developing genetically stable, high-yield strains is crucial for industrial-scale production.In this study, starting with Escherichia coli W3110 as the parent strain, a systematic metabolic engineering strategy was employed.This involved knocking out the L-tryptophan operon repressor gene (trpR) to relieve regulatory constraints, introducing feedback-resistant variants AroGfbr and TrpEfbr to enhance the flux of the L-tryptophan biosynthetic pathway, optimizing central metabolic pathways to increase the supply of precursors such as phosphoenolpyruvate (PEP) and erythrose-4-phosphate (E4P), and refining the L-tryptophan transport and cofactor supply systems.As a result, a high-yield strain SW-21 was constructed, which required no inducer, was plasmid-free, and exhibited genetic stability.The engineered strain SW-21, after 45 h of fed-batch fermentation in a 50 L fermenter, achieved an L-tryptophan titer of 57.2 g/L with a yield of 21.5%.This study integrated multiple metabolic engineering strategies to develop a strain suitable for industrial L-tryptophan production and provided a reference for constructing high-yield strains of other aromatic compounds.

Key words： Escherichia coli; L-tryptophan; adaptive evolution; systematic metabolic engineering; genome integration

Cite this article

WU Yadong , YANG Yan , WANG Jing , ZHANG Weifeng , WANG Xiaonan , CHEN Ning . Systematic metabolic engineering of Escherichia coli for high-yield L-tryptophan production[J]. Food and Fermentation Industries, 2026 , 52(14) : 48 -56 . DOI: 10.13995/j.cnki.11-1802/ts.044858

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