谷氨酸棒杆菌PhoPR双组分系统应答低氧胁迫功能研究  

Functional study of PhoPR two-component system in response to low oxygen stress in Corynebacterium glutamicum

作　　者：陈静 彭枫 刘秀霞[1,2,3] 杨艳坤 白仲虎[1,2,3] CHEN Jing;PENG Feng;LIU Xiuxia;YANG Yankun;BAI Zhonghu(National Engineering Laboratory of Cereal Fermentation Technology,Jiangnan University,Wuxi 214112,Jiangsu,China;Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,,Wuxi 214112,Jiangsu,China;School of Biotechnology,Jiangnan University,Wuxi 214112,Jiangsu,China)

机构地区：[1]江南大学粮食发酵工艺与技术国家工程试验室,江苏无锡214122 [2]江南大学工业生物技术教育部重点试验室,江苏无锡214122 [3]江南大学生物工程学院,江苏无锡214122

出　　处：《东北农业大学学报》2020年第5期24-31,共8页Journal of Northeast Agricultural University

基　　金：国家自然科学基金项目(21808082,21878124);江南大学研究生科研与实践创新计划项目(JNKY19_044)。

摘　　要：探究谷氨酸棒杆菌(Corynebacterium glutamicum,C. glutamicum)中PhoPR双组分系统响应外界溶氧变化功能机制。通过转录组数据分析发现PhoPR为C. glutamicum在不同溶氧水平下转录变化最明显双组分系统(Two-component system,TCS),表明PhoPR可感应环境中氧气变化。实时荧光定量PCR(Real-time quantitative PCR,RT-qPCR)和电泳迁移率试验(Electrophoretic mobility shift assay,EMSA)表明PhoPR效应蛋白PhoP可直接调控电子传递链相关基因sdhCAB和细胞膜相关基因betP、proP,间接调控qcrB,PhoPR高表达可增强细胞电子传递链和细胞膜基因转录。另外PhoPR还影响胞内ATP生成速度和峰值。基于以上结果推断,PhoPR通过调节电子传递链和ATP生成以维持C. glutamicum在低氧条件下生命活动。The mechanism of PhoPR two-component system(TCS) in response to the changes in dissolved oxygen was explored in Corynebacterium glutamicum(C. glutamicum). Transcriptome data analysis found that PhoPR was TCS with the most significant change of transcriptional levels under different dissolved oxygen conditions, indicating that PhoPR could sense changes of oxygen in the environment. Real-time quantitative PCR(RT-qPCR) and electrophoretic mobility shift assay(EMSA)showed that the response regulators(RR) protein PhoP in PhoPR could directly regulate the electron transfer chain related genes sdhCAB and cell membrane related genes betP and proP, and indirectly regulate qcrB. It was revealed that the overexpression of PhoPR could enhance the transcription of cel electron transfer chain and membrance genes. In addition, PhoPR affected the rate and peak of intracel ular ATP production. Based on the results, it was concluded that PhoPR regulates the electron transfer chain and ATP production to maintain the life activities of C. glutamicum under low oxygen condition.

关 键 词：谷氨酸棒杆菌 双组分系统 PhoPR 溶氧 EMSA 

分 类 号：Q815[生物学—生物工程]