文献类型：期刊文献

英文题名：Combining biosensor and metabolic network optimization strategies for enhanced l-threonine production in Escherichia coli

作者：Zhao, Zhenqiang[1,2];Zhu, Rongshuai[1,2];Shi, Xuanping[1,2];Yang, Fengyu[1,2];Xu, Meijuan[1,2];Shao, Minglong[1,2];Zhang, Rongzhen[1];Zhao, Youxi[3];You, Jiajia[1,2];Rao, Zhiming[1,2]

第一作者：Zhao, Zhenqiang

通讯作者：You, JJ[1];Rao, ZM[1];You, JJ[2];Rao, ZM[2]

机构：[1]Jiangnan Univ, Sch Biotechnol, Key Lab Ind Biotechnol, Minist Educ, Wuxi 214122, Jiangsu, Peoples R China;[2]JITRI, Inst Future Food Technol, Yixing 214200, Peoples R China;[3]Beijing Union Univ, Coll Biochem Engn, Beijing 100023, Peoples R China

第一机构：Jiangnan Univ, Sch Biotechnol, Key Lab Ind Biotechnol, Minist Educ, Wuxi 214122, Jiangsu, Peoples R China

通讯机构：[1]corresponding author), Jiangnan Univ, Sch Biotechnol, Key Lab Ind Biotechnol, Minist Educ, Wuxi 214122, Jiangsu, Peoples R China;[2]corresponding author), JITRI, Inst Future Food Technol, Yixing 214200, Peoples R China.

年份：2025

卷号：18

期号：1

外文期刊名：BIOTECHNOLOGY FOR BIOFUELS AND BIOPRODUCTS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001451654300001)】；

基金：The National Key Research and Development Program of China (2021YFC2100900). The National Natural Science Foundation of China (32071470). The National First-class Discipline Program of Light Industry Technology and Engineering (QGJC20230201). The Major Scientific and Technological Project for "unveiling and commanding" of Hohhot (2023-unveiling and commanding-He-1). The Open Funding Project of Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University (KLIB-KF202102). The Program of Introducing Talents of Discipline to Universities (111-2-06) and Postgraduate Research & Practice Innovation Program of Jiangsu Provence (KYCX22-2369).

语种：英文

外文关键词：l-Threonine biosensor; Directed evolution; High-throughput; Multi-omics analysis; In silico simulation

摘要：l-threonine is an integral nutrient for mammals, often used in animal feeds to enhance growth and reduce breeding costs. Developing l-threonine engineered strains that meet industrial production specifications has significant economic value. Here, we developed a biosensor that monitors l-threonine concentration to assist in high-throughput screening to capture high-yielding l-threonine mutants. Among them, the PcysK promoter and CysB protein were used to construct a primary l-threonine biosensor, and then the Cys(BT102A) mutant was obtained through directed evolution resulting in a 5.6-fold increase in the fluorescence responsiveness of biosensor over the 0-4 g/L l-threonine concentration range. In addition, the metabolic network of mutant was further optimized through multi-omics analysis and in silico simulation. Ultimately, the THRM13 strain produced 163.2 g/L l-threonine, with a yield of 0.603 g/g glucose in a 5 L bioreactor. The biosensor constructed here could be employed for iterative upgrading of subsequent strains, and these engineering strategies described provide guidance for other chemical overproducers.