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 1 ; Wang, Yuheng 1 ; Wang, Xiaolu 1 ; Bai, Yingguo 1 ; Wang, Yaru 1 ; Wang, Yuan 1 ; Tu, Tao 1 ; Qin, Xing 1 ; Su, Xiaoyun 1 ; Yao, Bin 1 ; Luo, Huiying 1 ; Liu, Xu 1 ; Huang, Huoqing 1 ; Zhang, Jie 1 ;
作者机构： 1.Chinese Acad Agr Sci, Inst Anim Sci, State Key Lab Anim Nutr & Feeding, Beijing 100193, Peoples R China
关键词： Escherichia coli; l-isoleucine; metabolic engineering; citramalate pathway; threonine-independent pathway
期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：6.2； 五年影响因子：6.4 ）
ISSN： 0021-8561
年卷期： 2025 年 73 卷 19 期
页码：
收录情况： SCI
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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