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巨大芽胞杆菌B196菌株抑菌物质的分离纯化

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摘要巨大芽胞杆菌(Bacillus megaterium)B196菌株产生的拮抗物质对水稻纹枯病菌(Rhizoctonia solani)的生长具有较强的抑制作用,明确该菌产生拮抗物质的种类是进一步研究该菌的抑菌机制及其应用的基础。本文采用盐酸沉淀B196菌株的去菌体培养液,再用甲醇抽提获得拮抗物质的粗提物。利用反相HPLC系统,将粗提物过C18柱,收集有抑制水稻纹枯病菌生长作用的活性化合物。运用质谱测得其分子量分别为1 042.592 7 u,再利用碰撞诱导解离(CID)技术获得化合物的典型结构特征离子碎片,结果表明其一级结构为ProAsnSerβAAAsnTyrAsnGln (βAA为14个碳原子的氨基脂肪酸)。綜合以上信息将该化合物鉴定为Iturin A2。

关键词巨大芽胞杆菌;iturin A2;分离纯化

中图分类号:S482.292文献标识码:ADOI:10.3969/j.issn.05291542.2014.02.004Isolation and purification of an antimicrobial substance produced

by Bacillus megaterium strain B196 Liao Ting,Qin Jian,Yuan Gaoqing,Li Qiqin,Lin Wei,Peng Haowen(College of Agriculture,Guangxi University, Nanning530005, China)AbstractAn antimicrobial substance produced by Bacillus megaterium strain B196 can strongly inhibit the growth of Rhizoctonia solani.Identification of the antimicrobial substance is very important for further research on the inhibition mechanism and its application.Crude antimicrobial substances were extracted with methanol from the precipitate which was obtained by adding HCl to the cellfree culture broth and then stored at 4 ℃ overnight.The crude extract was further purified by reversedphase HPLC with a C18 column(PRCODS,20×250 nm)to separate the substances.An antifungal compound with strong inhibitory activity against R.solani was purified.The molecular weight of the compound was 1 042.592 7 u by Matrixassisted laser desorption ionization timeofflight mass spectrometer(MALDITOFMS).TESI collision induced dissociation mass spectrometry analysis demonstrated that the compou...



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