寡养单胞菌膜孔蛋白D0Y85_RS06780对细菌抗生素耐药性的作用研究  

Study on the effect of cell membrane porin D0Y85_RS06780 of Stenotrophomonas sp.on bacterial antibiotic resistance

作　　者：刘亚豫 任钊锐 谢欣茹 王祯 王高扬 曹广祥 付加芳 Liu Ya-yu;Ren Zhao-rui;Xie Xin-ru;Wang Zhen;Wang Gao-yang;Cao Guang-xiang;Fu Jia-fang(College of Biomedical Sciences,Shandong First Medical University&Shandong Academy of Medical Sciences,Jinan 250117;School of Municipal and Environmental Engineering,Shandong Jianzhu University,Jinan 250101)

机构地区：[1]山东第一医科大学(山东省医学科学院)生物医学科学学院,济南250117 [2]山东建筑大学市政与环境工程学院,济南250101

出　　处：《中国抗生素杂志》2023年第4期426-432,共7页Chinese Journal of Antibiotics

基　　金：山东省自然科学基金项目(No.ZR2022MH107);山东第一医科大学学术提升计划(No.2019 LJ001)。

摘　　要：目的探究寡养单胞菌Stenotrophomonas sp.G4菌株中膜孔蛋白D0Y85_RS06780对抗生素耐药性的作用。方法通过氨基酸序列比对和同源建模预测D0Y85_RS06780蛋白的生物学功能;采用大肠埃希菌构建D0Y85_RS06780基因的异源表达菌株,通过检测抗生素MICs探讨D0Y85_RS06780对细菌抗生素抗性的作用,同时分析外排泵抑制剂对D0Y85_RS06780功能的影响;利用分子对接软件DiscoveryStudio的CDOCKER技术分析D0Y85_RS06780与抗生素和外排泵抑制剂的结合情况。结果同源分析表明D0Y85_RS06780为膜孔蛋白,异源表达D0Y85_RS06780导致重组大肠埃希菌具有多黏菌素和四环素抗性,外排泵抑制剂利血平能够抑制其功能,分子对接显示D0Y85_RS06780能结合多黏菌素、四环素和利血平。结论Stenotrophomonas sp.G4菌株中膜孔蛋白D0Y85_RS06780与其抗生素耐药性有关。本研究将为D0Y85_RS06780与抗生素耐药性的关联提供实验证据。Objective To explore the effect of cell membrane porin D0Y85_RS06780 of Stenotrophomonas sp.G4 on bacterial antibiotic resistance.Methods Biological function of D0Y85_RS06780 was predicted based on amino acid sequence alignment and homologous modeling;D0Y85_RS06780 gene was heterologously expressed in Escherichia coli,and its effect on bacterial antibiotic resistance was determined through MICs;The effect of efflux pump inhibitors on D0Y85_RS06780 were also analyzed through antibiotic MICs;The molecular docking was carried out according to the CDOCKER protocol of Discovery Studio to determine whether D0Y85_RS06780 can bind antibiotics and efflux pump inhibitors.Results Homologous analysis showed that D0Y85_RS06780 is a membrane porin protein.Recombinant E.coli expressing D0Y85_RS06780 obtained polymyxin and tetracycline resistance,and the polymyxin resistance could be inhibited by the efflux pump inhibitor reserpine.Molecular docking revealed that D0Y85_RS06780 could bind polymyxin,tetracycline and the efflux pump inhibitor reserpine.Conclusion Cell membrane porin D0Y85_RS06780 of Stenotrophomonas sp.is associated with bacterial antibiotic resistance.This study provides experimental evidence for further research on antibiotic resistance mechanism of cell membrane porin D0Y85_RS06780.

关 键 词：寡养单胞菌 膜孔蛋白 耐药性 异源表达 分子对接 

分 类 号：R917[医药卫生—药物分析学]