人肠道微生物中抗菌活性菌株的筛选及其代谢产物研究
投稿时间:2019-06-11     点此下载全文
引用本文:李德利,丁鹏敏,刘双月,杨秀伟,王如峰.人肠道微生物中抗菌活性菌株的筛选及其代谢产物研究[J].中国现代中药,2020,22(1):26-34
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作者中文名作者英文名单位中文名单位英文名E-Mail
李德利 LI De-li 北京中医药大学 生命科学学院,北京102488 School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488,China  
丁鹏敏 DING Peng-min 北京中医药大学 生命科学学院,北京102488 School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488,China  
刘双月 LIU Shuang-yue 北京中医药大学 生命科学学院,北京102488 School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488,China  
杨秀伟 YANG Xiu-wei 北京大学 药学院,北京100083 School of Pharmaceutical Sciences,Peking University,Beijing 100083,China  
王如峰 WANG Ru-feng 北京中医药大学 生命科学学院,北京102488 School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488,China 王如峰,教授,研究方向:中药化学;Tel:(010)84738646,E-mail:wangrufeng@tsinghua.org.cn 
基金项目:国家重点研发计划(2018YFC1704500)
中文摘要:目的:从肠道菌群中筛选抗菌活性菌株,明确其抗菌活性物质,为认识肠道微生物之间的相互作用乃至与宿主健康的关系奠定基础。方法:对肠道混合菌群进行体外培养,以抗菌活性为指标筛选活性单菌株;采用生化鉴定法和16S rDNA鉴定法对菌株进行鉴定;通过活性追踪手段,运用各种色谱分离技术对菌株的代谢产物进行分离纯化,并利用光谱技术鉴定其结构;采用微量孔板法测定各化合物的最低抑菌浓度(MIC)。结果:筛选得到具有抗菌作用的普通变形杆菌Proteus vulgaris HA-3151菌株;从其发酵液中分离得到13个化合物:D-3-苯基乳酸、环(丙-苯丙)二肽、环(酪-酪)二肽、环(酪-亮)二肽、环(酪-异亮)二肽、丁香酸、黄豆黄苷、大豆苷、大豆苷元、染料木苷、5-甲氧基大豆苷元、3-甲氧基没食子酸和染料木黄酮;其中,D-3-苯基乳酸、大豆苷元、染料木苷、染料木黄酮以及环(丙-苯丙)二肽、环(酪-酪)二肽具有抑菌作用。结论:首次得到了有抗菌活性的普通变形杆菌Proteus vulgaris菌株并证明其可产生有机酸、黄酮和二肽类抗菌活性物质,这些物质有助于其形成定植优势,进而对人体健康产生各种影响。
中文关键词:肠道微生物  普通变形杆菌  代谢产物  抗菌活性
 
Screening of An Antibacterial Strain from Human Intestinal Microbes and Investigation on Its Active Metabolites
Abstract:Objective:To screen antibacterial strain from human intestinal microbes and ascertain its antibacterial active substances,so as to lay a foundation for understanding the interaction between intestinal microorganisms and its relationship with host health.Methods:Intestinal microbiota was cultured in vitro,and the active strain was screened using antibacterial activity as an index.The strain was identified by biochemical test and 16S rDNA sequencing.The metabolites of the strain were isolated and purified through activity tracking using various chromatographic separation techniques,and their structures were identified on the basis of spectral analysis.The minimum inhibitory concentration (MIC) of each compound was determined by microplate method.Results:A strain named Proteus vulgaris HA-3151 with antibacterial activity was screened out,and 13 compounds including D-3-phenyllactic acid,cycle(Ala-Phe),cycle(Tyr-Tyr),cycle(Tyr-Leu),cycle(Tyr-Ile),syringic acid,glycitin,daidzin,daidzein,genistin,5-methoxydaidzein,3-methoxygallic acid,and genistein were isolated from its fermentation broth.Among them,6 compounds including D-3-phenyllactic acid,cycle(Ala-Phe),cycle(Tyr-Tyr),daidzein,genistin,and genistein showed antibacterial activity.Conclusion:The Proteus vulgaris strain with antibacterial activity was obtained for the first time and it was proved to produce antibacterial active substances such as organic acids,flavonoids and dipeptides,and these substances may contribute to the colonization advantage and further affect human health.
keywords:intestinal microbes  Proteus vulgaris  metabolites  antibacterial activity
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