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强化hemE和hemQ表达对枯草芽孢杆菌合成血红素的影响 认领

Effect of overexpressing hemE and hemQ on heme synthesis in Bacillus subtilis
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摘要 枯草芽孢杆菌(Bacillus subtilis)作为公认安全(Generally Recognized as Safe,GRAS)菌株,是生产食品级血红素的理想底盘,但其粪卟啉依赖性(coproporphyrin-de...展开更多 Bacillus subtilis,a Generally Recognized as Safe(GRAS)strain,serves as an ideal chassis for producing food-grade heme.However,the regulatory mechanisms of key enzymes within its coproporphyrin-dependent(CPD)synthesis pathway remain unclear.To elucidate and overcome the ...MORE Bacillus subtilis,a Generally Recognized as Safe(GRAS)strain,serves as an ideal chassis for producing food-grade heme.However,the regulatory mechanisms of key enzymes within its coproporphyrin-dependent(CPD)synthesis pathway remain unclear.To elucidate and overcome the rate-limiting steps in this pathway,this study systematically evaluated the impact of enhancing the expression of uroporphyrinogen decarboxylase(UroD,encoded by hemE)and coproheme decarboxylase(ChdC,encoded by hemQ)on heme synthesis.The genes hemE and hemQ,controlled by a strong constitutive promoter,were integrated into the neutral genomic loci ydaP and yvkC of B.subtilis BSH11,both individually and in combination,using a markerless gene modification technique.The resulting engineered strains were then evaluated for heme production in shake-flask fermentation and for gene transcription levels by Real-time quantitative PCR(RT-qPCR).The results showed that overexpressing hemE alone yielded the most significant improvement.Its transcription level increased by up to 303-fold,leading to an extracellular heme production of 23.90±1.49 mg/L after 48 h of fermentation,an 826%increase compared to the control strain.In contrast,overexpression of hemQ alone did not significantly promote heme production.Furthermore,the co-expression of both genes resulted in a 35%decrease in extracellular heme yield compared to the strain overexpressing only hemE.These findings demonstrate that UroD is a key rate-limiting step in the CPD pathway of B.subtilis,whereas the catalytic ability of ChdC is not the primary limiting factor under the current conditions.This discovery identifies a core target and provides theoretical guidance for efficiently synthesizing food-grade heme in the safe B.subtilis chassis via precise metabolic engineering.FEWER
作者 杨绍梅 周长旭 王鹏 张同 来凤堂 李树标 薄文文 马钦元 YANG Shaomei;ZHOU Changxu;WANG Peng;ZHANG Tong;LAI Fengtang;LI Shubiao;BO Wenwen;MA Qinyuan(Jiangsu Shenhua Pharmaceutical Co.Ltd.,Huai’an 211600,China;School of Life Sciences and Medicine,Shandong University of Technology,Zibo 255000,China;Shandong Fufeng Fermentation Co.Ltd.,Linyi 276000,China)
机构地区 江苏神华药业有限公司 山东理工大学,生命与医药学院 山东阜丰发酵有限公司
出处 《食品与发酵工业》 EI CAS CSCD 北大核心 2026年第12期35-42,共8页 Food and Fermentation Industries
基金 山东省自然科学基金项目(ZR2022QC078)。
关键词 血红素 枯草芽孢杆菌 代谢工程 hemE基因 基因过表达 heme Bacillus subtilis metabolic engineering hemE gene gene overexpression
分类号 TS201.3 [轻工技术与工程—食品科学]
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食品与发酵工业
2026年 第12期
科创助手
论文智评
维普砚承
维普斟知

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