数字农科院2.0

Metabolic Engineering of Escherichia coli for Efficient l-Isoleucine Production based on the Citramalate Pathway

文献类型： 外文期刊

作者： Zhang, Qiquan;Wang, Yuheng;Wang, Xiaolu;Bai, Yingguo;Wang, Yaru;Wang, Yuan;Tu, Tao;Qin, Xing;Su, Xiaoyun;Yao, Bin;Luo, Huiying;Liu, Xu;Huang, Huoqing;Zhang, Jie

作者机构：

关键词： Escherichia coli;l-isoleucine;metabolic engineering;citramalate pathway;threonine-independent pathway

期刊名称： JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY

ISSN： 0021-8561

年卷期： 2025 年

页码：

收录情况： SCIE（2025版） ; ; EI（2025版）

摘要： l-isoleucine, a value-added branched-chain amino acid, has been widely used in the food, feed, and pharmaceutical industries. However, the production efficiency of l-isoleucine is relatively low due to the complex and inefficient biosynthetic network of the canonical threonine pathway. Here, we report the exploitation of a concise citramalate pathway for the development of an efficient l-isoleucine producer. First, chassis strain and key genes were screened for establishment of the citramalate pathway. Subsequently, a citramalate importer was identified and applied to enhance citramalate's utilization efficiency. Finally, a plasmid-free high-l-isoleucine producer was developed by enhancement of l-isoleucine efflux, optimization of rate-limiting gene expression, and introduction of a nonoxidative glycolysis pathway. Fed-batch fermentation of the final strain in a 10-L bioreactor produced 56.6 g/L l-isoleucine with a productivity of 1.66 g/L/h, which is the highest l-isoleucine titer and productivity reported. This study paves the way for construction of efficient microbial cell factories for production of l-isoleucine and related derivatives based on the citramalate pathway.

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