文献类型：期刊文献

英文题名：Metabolic engineering of Escherichia coli for squalene overproduction

作者：Zhu, Jiangming[1,2,3];Mao, Yaping[1,2,3];Mo, Hongchun[2,3];Dai, Xuehui[1,2,3];Wu, Yuhan[1,2,3];Wang, Guangyi[1,2,3];Feng, Zhanguang[1,2,3];Yue, Ruirui[1,2,4];Wei, Dongzhi;Liu, Haili[1,2];Wang, Yong[1,2,3]

机构：[1]East China Univ Sci & Technol, Sch Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Chinese Acad Sci, CAS Ctr Excellence Mol Plant Sci, CAS Key Lab Synthet Biol, Shanghai 200032, Peoples R China;[3]Univ Chinese Acad Sci, Beijing 100039, Peoples R China;[4]Henan Univ, Sch Life Sci, Kaifeng 475004, Peoples R China

年份：2025

卷号：10

期号：4

起止页码：1119

外文期刊名：SYNTHETIC AND SYSTEMS BIOTECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001533947500001)】；

基金：This work is financially supported by the National Key R & D Program of China (No. 2024YFA0919900) , the Key Laboratory of Shanghai Tobacco Group Cigarette Smoke (No. K2023-1-044P) , the State Key Laboratory of Plant Trait Design, the Key Laboratory of Plant Carbon Capture, Chinese Academy of Sciences, and the Shanghai Municipal Science and Technology Major Project. We thank Dr. Sheng Yang (Key Laboratory of Synthetic Biology, CAS Principal Investigator) for providing the plasmids pEccas and pTarget used in E. coli genome editing.

语种：英文

外文关键词：Squalene; Escherichia coli; Metabolic engineering; Targeted lipidomics

摘要：Squalene, a lipophilic triterpene with multifaceted bioactivities, faces bioproduction bottlenecks in microbial hosts due to inefficient biosynthetic pathways and limited storage capacity. Here, we address these challenges through systems metabolic engineering integrating redox-balanced 3-hydroxy-3-methyl glutaryl coenzyme A reductase (HMGR) variants and membrane lipid remodeling. By developing a hybrid HMGRs system combining NADPH-dependent and NADH-preferred enzymes, squalene production reached 852.06 +/- 28.95 mg/L with balanced cofactor utilization. Subsequent engineering of membrane morphology and lipid metabolism generated lipid-enriched elongated cells, through the overexpression of dgs, murG and plsC, boosting squalene production to 970.86 +/- 55.67 mg/L. Implementation of delayed induction strategies coupled with 10 % dodecane overlay as an in situ recovery system achieved a final squalene titer of 1267.01 mg/L in a 3 L bioreactor. Mechanistic studies revealed fatty acid (FA) and phosphatidylethanolamine (PE) as key reservoirs for squalene in E. coli, with dgs overexpression specifically promoting cellular elongation. This article provides comprehensive insights into engineering strategies and mechanistic perspectives, establishing a universal framework for hydrophobic metabolite biomanufacturing in prokaryotic hosts.