食品与发酵工业 >

2024 , Vol. 50 >Issue 15: 1 - 7

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.039589

研究报告

蓝光调控生物膜关键基因提高大肠杆菌耐酸性

  • 吴建菊 ,
  • 冯守帅
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  • (江南大学 生物工程学院,工业生物技术教育部重点实验室,江苏 无锡,214122)
第一作者：硕士研究生(冯守帅教授为通信作者,E-mail:fengss@jiangnan.edu.cn)

收稿日期: 2024-04-17

  修回日期: 2024-04-29

  网络出版日期: 2024-08-21

基金资助

国家自然科学基金项目(21878128)

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Enhancing the acid resistance of Escherichia coli by blue light-regulated biofilm key genes

  • WU Jianju ,
  • FENG Shoushuai
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  • (The Key Laboratory of Industrial Biotechnology, Ministry of Educations, School of Biotechnology, Jiangnan University, Wuxi 214122, China)

Received date: 2024-04-17

  Revised date: 2024-04-29

  Online published: 2024-08-21

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摘要

发酵过程中的酸胁迫限制了工业微生物的发展,精细调控生物膜形成是提升大肠杆菌耐酸性的有效策略。研究基于EL222蓝光诱导蛋白开发了具有8.6倍调控效率的双质粒调控系统,用于调节生物膜形成关键因子CsgD和二鸟苷酸环化酶M(diguanylate cyclase,DgcM)的表达。结果表明,在pH 5.0和15 mW/cm2蓝光照射条件下,CsgD和DgcM过表达菌株耐酸性分别提高了22.3%和16.4%,生物膜含量分别增加了2.7倍和3.4倍。生物膜组成分析显示,酸性环境下多糖和蛋白质的增加以及可溶性微生物代谢产物的减少促进了细菌聚集,有助于形成更稳定的生物膜结构。实时荧光定量逆转录PCR结果证实生物膜形成相关基因转录水平显著提升。本研究构建的蓝光调控系统通过调控生物膜形成关键基因成功提高了大肠杆菌的耐酸性,这将为微生物的有机酸发酵提供参考和借鉴。

关键词： 大肠杆菌; 蓝光调控; 生物膜; 耐酸性; 实时荧光定量逆转录PCR(RT-qPCR)

本文引用格式

吴建菊 , 冯守帅 . 蓝光调控生物膜关键基因提高大肠杆菌耐酸性[J]. 食品与发酵工业, 2024 , 50(15) : 1 -7 . DOI: 10.13995/j.cnki.11-1802/ts.039589

Abstract

Acidic stress has limited the development of industrial microorganisms during the fermentation process.Fine regulation of biofilm formation is an effective strategy to improve the acid tolerance of Escherichia coli.A dual-plasmid regulatory system based on the EL222 blue light-inducible protein was successfully constructed with an 8.6-fold increase in regulatory efficiency.This dual-plasmid expression system was further used to regulate the expressions of a biofilm formation key regulatory factor CsgD and diguanylate cyclase M (DgcM) proteins.The results indicated that the acid resistance of CsgD and DgcM overexpressed strains was improved by 22.3% and 16.4% under conditions of pH 5.0 and 15 mW/cm2 blue light irradiation.Moreover, the biofilm content was increased by 2.7 and 3.4 times, respectively.The analysis of biofilm composition revealed that the level of polysaccharides and proteins increased, while the soluble microbial products decreased under acid stress.These changes promoted bacterial aggregation, contributing to the formation of a more stable biofilm structure.Quantitative reverse transcription PCR (RT-qPCR) results further showed a significant upregulation in the transcription levels of biofilm formation-related genes.This blue light-regulated system successfully improved the acid resistance of E.coli by controlling key genes for biofilm formation, which will offer potential references and guidance for microbial organic acid fermentation.

Key words： Escherichia coli; blue light-regulated; biofilm; acid resistance; quantitative reverse transcription PCR (RT-qPCR)

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