甲基营养菌Methylorubrum extorquens AM1抗菌活性物质分离与鉴定  

Isolation and Identification of Antibacterial Activity Substances from Methylorubrum extorquens AM1

作　　者：李婉婷 鲍梦姣 张聪 杨松[1] 于桂洪 LI Wanting;BAO Mengjiao;ZHANG Cong;YANG Song;YU Guihong(Shandong Province Key Laboratory of Applied Mycology/School of Life Science,Qingdao Agricultural University,Qingdao 266109,China)

机构地区：[1]青岛农业大学生命科学学院/山东省应用真菌重点实验室,山东青岛266109

出　　处：《青岛农业大学学报（自然科学版）》2023年第2期124-131,共8页Journal of Qingdao Agricultural University(Natural Science)

基　　金：青岛农业大学高层次人才项目(665/1120034);山东省重点研发计划(2021ZDSYS28)。

摘　　要：甲基营养菌能够以一碳化合物作为唯一碳源和能源进行生长,培养成本较低,已用来生产多种高附加值产品和药用活性物质。因此,挖掘甲基营养菌的抗菌代谢产物,将为抗菌物质的研发和生产提供研究基础。本研究通过肉汤微量稀释法进行抗菌活性初筛,利用7.5 L发酵罐对活性菌株Methylorubrum extorquens AM1进行放大发酵,采用硅胶柱层析和高效液相色谱等对粗提物进行分离纯化,应用核磁共振、质谱、旋光等手段确定化合物的化学结构,共得到7种化合物,分别为腺苷(1)、5′-S-methyl-5′-thioadenosine(2)、肌苷(3)、尿苷(4)、1,1′-oxybis(2,4-di-tert-butylbenzene)(5)、胸腺嘧啶(6)和烟酰胺(7),其中,化合物1至5为首次在Methylorubrum属的甲基营养菌中分离得到。抗菌活性评价显示化合物1至4对枯草芽孢杆菌、志贺菌和/或金黄杆菌具有一定抑制作用,MIC 50在2.1~375.0μg/mL之间。Methylotrophic bacteria are capable of using one-carbon compounds as the sole carbon and energy sources for growth,with low cultivation costs.They have been utilized to produce a variety of high value-added products and medicinal substances.Therefore,exploring antimicrobial metabolites from methylotrophic bacteria provides a research foundation for developing and producing antimicrobial agents.In this study,broth microdilution method was used to measure the antibacterial activity of crude extracts from methylotrophic bacteria.Scaled up fermentation of the bioactive strain Methylorubrum extorquens AM1 was carried out using a 7.5 L fermenter.The resulting total crude extract was separated and purified by silica gel column chromatography and high performance liquid chromatography(HPLC).The structures of metabolites were determined by means of nuclear magnetic resonance(NMR),mass spectrometry(MS),optical rotation(OR),et al.As a result,7 compounds were obtained,which were determined as adenosine(1),5′-S-methyl-5′-thioadenosine(2),inosine(3),uridine(4),1,1′-oxybis(2,4-di-tert-butylbenzene)(5),thymine(6)and nicotinamide(7).Compounds 1 to 5 were isolated from Methylorubrum for the first time.Activity evaluation showed that compounds 1 to 4 inhibited the growth of Bacillus subtilis,Shigella sp.and/or Chryseobacterium sp.,with MIC 50 of 2.1-375.0μg/mL.

关 键 词：甲基营养菌 Methylorubrum extorquens 代谢产物 抗菌活性 

分 类 号：Q939.92[生物学—微生物学]