食品与发酵工业2025，Vol.51Issue(12)：1-10,10.DOI:10.13995/j.cnki.11-1802/ts.040184
双向启动子的开发及其在大肠杆菌甲羟戊酸生产中的应用
Development of bidirectional promoters and their applications to production of mevalonate in Escherichia coli
摘要
Abstract
Bidirectional promoters(BDPs)are defined as promoters possessing an opposing transcription site within 7-25 bp of both the right and left ends of the transcription start site of the promoter.BDPs serve as gene elements that coordinate the expression of different genes,and can lead to the inactivation of both genes if a single side of the element is compromised,thereby facilitating co-regulation.However,there is a lack of efficient and rapid methods for screening highly active BDPs in prokaryotes.In this study,a method was devel-oped to rapidly screen for highly active BDPs in prokaryotes.Based on previous laboratory work,the active probe vector p19BDP for rapid BDP screening on plates was developed.Finally,two highly active BDPs from Escherichia coli and one from Corynebacterium glutamicum,each with heterozygous-10 or-35 regions,were identified.Subsequently,seven symmetrical libraries were developed to identify eight highly active BDPs exhibiting bilateral activity in C.glutamicum.Fluorescence detection confirmed that these eight promoters also func-tioned bidirectionally in E.coli.The selected BDPs were utilized for the production of mevalonate(MVA)in E.coli.Compared to the T7 promoter induced with 0.5 mmol/L isopropyl-beta-D-thiogalactopyranoside,BDP-W6 expressing genes on both sides achieved a similar MVA yield to the control strain at 24 hours(19.8 mg/L).At 48 hours,the MVA yield reached 72.6%of the control strain's yield,to-talling 32.2 mg/L.This study outlines a research methodology for the large-scale screening of BDPs and confirms their effectiveness.Com-pared with the induced expression of the T7 promoter,the bidirectional high-activity expression of BDPs is more economical.关键词
大肠杆菌/谷氨酸棒杆菌/双向启动子/甲羟戊酸/探针载体Key words
Escherichia coli/Corynebacterium glutamicum/bidirectional promoters/mevalonate/probe vector引用本文复制引用
段元朔,付振浩,刘秀霞,白仲虎..双向启动子的开发及其在大肠杆菌甲羟戊酸生产中的应用[J].食品与发酵工业,2025,51(12):1-10,10.基金项目
国家自然科学基金重点项目(21938004) （21938004）
