生物加工过程2025，Vol.23Issue(2)：190-198,9.DOI:10.3969/j.issn.1672-3678.2025.02.009
产长叶烯大肠杆菌工程菌的构建及发酵优化
Construction of engineered Escherichia coli to produce longlifolene and optimization of the fermentation process
摘要
Abstract
In this study,Escherichia coli(E.coli)BL21(DE3)was used as the chassis cell.Farnesyl pyrophosphate(FPP)synthase from different sources were screened and key enzyme genes in the endogenous MEP pathway,including 1-deoxy-D-xylulose-5-phosphate synthase(dxs),1-deoxy-D-xylulose-5-phosphate reductoisomerase(dxr),isopentenyl pyrophosphate isomerase(idi)were overexpressed.An engineered E.coli cell factory with an optimized central carbon metabolic pathway capable of synthesizing longifolene efficiently was constructed.To further enhance the precursor supply for the synthesis of longleafene in the cell factory,fermentation conditions were optimized in this study.It was indicated that the optimal concentration of the inducer isopropyl β-D-thiogalactoside(IPTG)was 0.1 mmol/L,with the preferable addition time being 0 h(when the seed culture was transferred to the fermentation broth)and the optimal induction temperature at 30 ℃.The most suitable medium was TB medium.Finally,the yield of longifolene in the shake flask reached 3.85 mg/L,a 45.8％increase compared to the value reported in the literature.This study laid a good foundation for further research to realize the high yield of longifolene using E.coli.关键词
MEP途径/长叶烯/大肠杆菌/发酵优化Key words
MEP pathway/longifolene/Escherichia coli/fermentation optimization分类
生物学引用本文复制引用
夏芙蓉,魏丽娟,刘欢,巴利民,刘艳辉..产长叶烯大肠杆菌工程菌的构建及发酵优化[J].生物加工过程,2025,23(2):190-198,9.基金项目
国家自然科学基金青年项目(21606014) （21606014）
