食品与发酵工业 >

2026 , Vol. 52 >Issue 1: 18 - 24

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.042384

研究报告

构建大肠杆菌C4途径强化血红素合成

  • 王子晨 ,
  • 张鑫 ,
  • 唐蕾
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  • 1(江南大学 生物工程学院,江苏 无锡,214122)
    2(工业生物技术教育部重点实验室(江南大学),江苏 无锡,214122)
第一作者:硕士研究生(唐蕾教授为通信作者,E-mail:ltang@jiangnan.edu.cn)

收稿日期: 2025-02-24

  修回日期: 2025-03-25

  网络出版日期: 2026-01-22

基金资助

111引智计划项目(111-2-06);国家轻工技术与工程一流学科自主课题资助项目(LITE2018-27)

收起

Enhancing heme biosynthesis in Escherichia coli by constructing C4 pathway

  • WANG Zichen ,
  • ZHANG Xin ,
  • TANG Lei
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  • 1(School of Biotechnology, Jiangnan University, Wuxi 214122, China)
    2(Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China)

Received date: 2025-02-24

  Revised date: 2025-03-25

  Online published: 2026-01-22

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摘要

血红素一般指亚铁血红素,由亚铁离子和原卟啉组成,是铁卟啉的衍生物,目前在食品医疗等领域的需求日益增加。微生物发酵是获取血红素的方法之一,但是产量低。该研究以大肠杆菌(Escherichia coli)BL21(DE3)为底盘菌株,通过代谢工程手段改造大肠杆菌,获得了高产血红素的基因工程菌株。首先阻断了大肠杆菌原有的血红素合成C5途径,导入异源的C4途径,获得的基因工程菌血红素产量优于强化原有C5途径或C4/C5途径共存的菌株。继而在该工程菌株中过表达大肠杆菌血红素合成途径的关键基因hemB或hemH,血红素的产量进一步提升。最终通过优化前体甘氨酸和FeCl2的供给,经过24 h摇瓶发酵,血红素产量达到了82.39 μmol/L,是原始出发菌株的4.2倍。该研究构建了仅表达异源C4途径的大肠杆菌菌株,为代谢工程改造提高大肠杆菌血红素的合成提供了新的策略。

关键词： 血红素; 大肠杆菌; C4途径; C5途径; 代谢工程

本文引用格式

王子晨 , 张鑫 , 唐蕾 . 构建大肠杆菌C4途径强化血红素合成[J]. 食品与发酵工业, 2026 , 52(1) : 18 -24 . DOI: 10.13995/j.cnki.11-1802/ts.042384

Abstract

Heme is an iron porphyrin derivative composed of a ferrous ion and protoporphyrin.The demand of heme is increasing in fields such as food and medicine.Microbial fermentation is one of the methods for obtaining heme, but the titer is low.In this study, Escherichia coli BL21(DE3) was used as chassis strain and genetically modified to obtain a high-yield heme producing strain through metabolic engineering strategies.Firstly, the native C5 heme biosynthetic pathway in E.coli was blocked and a heterologous C4 pathway was introduced.The resulting engineered strain demonstrated superior heme production compared to strains with enhanced native C5 pathway or coexisting C4/C5 pathways.Subsequently, a further increase of heme production was achieved by over expressing the key genes of hemB or hemH involved in the heme synthetic pathway in E.coli.Finally, by optimizing the supplement of the precursors of glycine and ferrous chloride, the heme content in the engineered strain reached 82.39 μmol/L after 24 h of shaking flask fermentation, which was 4.2-fold compared to that of the starting strain.It is a new strategy to construct an engineered E.coli which uses sole heterologous C4 pathway to produce heme with high titer.

Key words： heme; E.coli; C4 pathway; C5 pathway; metabolic engineering

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