基于蛋白组学对苯胺降解菌Rhodococcus sp.AN-P1苯胺胁迫响应的研究  

Research on Aniline-degrading Bacterium Rhodococcus sp.AN-P1 in Response to Aniline Based on Proteomics

作　　者：刘冰霜[1] 王臣 杨冲 刘庆华[1] 谭周亮[1] LIU Bingshuang;WANG Chen;YANG Chong;LIU Qinghua;TAN Zhouliang(Chengdu Institute of Biology,Chinese Academy of Sciences/Key Laboratory of Environmental and Applied Microbiology,Chinese Academy of Sciences,Chengdu 610041,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院成都生物研究所/中国科学院环境与应用微生物重点实验室,四川成都610041 [2]中国科学院大学,北京100049

出　　处：《广东农业科学》2020年第4期85-91,共7页Guangdong Agricultural Sciences

基　　金：四川省重大科技专项(2019YFS0501);中国科学院青年创新促进会项目(2016331)。

摘　　要：【目的】研究微生物在苯胺胁迫环境下的响应机制,揭示Rhodococcus sp.AN-P1适应、降解苯胺内在分子机理,弥补苯胺降解菌在苯胺胁迫下耐受机制的空白,为进一步调控提高微生物降解苯胺能力提供良好的靶点,为其生物修复研究奠定理论基础。【方法】采用双向电泳技术分离纯化Rhodococcus sp.AN-P1在苯胺、柠檬酸条件下表达的蛋白质组,并利用基质辅助激光解析电离串联飞行时间质谱(MALDI-TOF-MS)鉴定显著差异表达蛋白,对比NCBInr蛋白数据库获得蛋白质点的详细信息。【结果】在以苯胺、柠檬酸为唯一碳源的条件下,Rhodococcus sp.AN-P1分别表达了681、579个蛋白质点;选取21个显著差异蛋白质点进行质谱分析,成功获得17个蛋白质点的相关信息,其分布于信号转导调节、氨基酸及能量代谢、细胞防御、苯胺降解酶多个代谢系统,并发现大部分蛋白质分子质量在31000~58000之间,等电点位于4~7之间。【结论】Rhodococcus sp.AN-P1在苯胺胁迫环境中会通过响应信号传导系统感知外界胁迫、调节氨基酸及能量代谢系统抵抗苯胺的毒害影响、利用细胞防御系统进一步提高其存活能力、表达苯胺降解酶系统降解苯胺获得碳源、氮源及能量来源,从而达到适应并降解高浓度苯胺的目的。【Objectives】The response mechanisms of microorganisms under aniline stress were studied to reveal the internal molecular mechanism for Rhodococcus sp.AN-P1 to adapt to and degrade aniline,supplement the gap of aniline-degrading bacterium tolerance mechanism under aniline stresss,provide a good target for further regulation and improvement of the ability of microorganisms to degrade aniline,and lay a theoretical foundation for the research of on bioremediation.【Methods】Two-dimensional electrophoresis was used to separate and purify the proteome of Rhodococcus sp.AN-P1 expressed under the conditions of aniline and citric acid.Matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry(MALDI-TOF-MS)was used to identify the significantly differentially expressed proteins.Finally,the detailed information of protein spots was obtained from the NCBInr protein database.【Results】With aniline and citric acid as the sole carbon source,Rhodococcus sp.AN-P1 expressed 681 and 579 protein spots,respectively.21 significantly different protein spots were selected for mass spectrometry analysis and related information of 17 protein spots was successfully obtained.These proteins were distributed in multiple metabolic systems such as signal transduction regulation,amino acid and energy metabolism,cell defense,and aniline degrading enzymes.It was found that the mass of most protein molecules was between 31000 and 58000,and the isoelectric point between 4 and 7.【Conclusions】Uunder the environment of aniline stress environment,Rhodococcus sp.AN-P1 could sense external stress by the signal transduction system under aniline stress environment,adjust the amino acid and energy metabolism system to resist the toxic effects of aniline,use the cell defense system to further improve its survival ability,and express aniline-degrading enzymes to degrade aniline for obtaining carbon source,nitrogen source and energy source.Ultimately,Rhodococcus sp.AN-P1 achieved the purpose of adapting to and degrading hig

关 键 词：蛋白组学 苯胺 胁迫响应 Rhodococcus sp.AN-P1 

分 类 号：Q933[生物学—微生物学]