摘要
【目的】建立一种从枯草芽胞杆菌中高效快速制备4种脂肽类抗生素的方法,初步阐明制备脂肽类抗生素的生物学活性。【方法】通过构建脂肽类抗生素突变株,采用酸沉淀、有机溶剂抽提和固相萃取的方法从枯草芽胞杆菌Bs916及其突变株的发酵液中提纯表面活性素、杆菌霉素L、泛革素和罗克霉素4种脂肽类抗生素;运用HPLC-MS、SDS-PAGE电泳对制备脂肽类抗生素的纯度进行检测;最后采用西瓜枯萎病菌、金黄色葡萄球菌和绵羊血红细胞测定4种脂肽类抗生素的抗真菌、抗细菌和溶血活性。【结果】蛋白凝胶电泳和液相色谱联用质谱检测结果表明,制备的脂肽类抗生素均达到了电泳纯和色谱纯;生物活性分析实验结果表明,表面活性素和杆菌霉素L具有较强的溶血活性,泛革素和罗克霉素的溶血活性较弱,杆菌霉素L和泛革素具有较强的抗真菌活性,而表面活性素和罗克霉素具有较强的抗细菌活性。【结论】建立快速从芽胞杆菌中提纯4种脂肽类抗生素色谱方法,测定了4种脂肽类化合物的生物活性,为进一步探究脂肽类抗生素的构效关系、生物合成和开发应用奠定基础。
[ Objective ] The efficient and simple methods for the preparation of four lipopeptide antibiotics produced by Bacillus subtilis strains were established and the biological functions of these four lipopeptide antibiotics were elucideted. [ Method] The four family lipopeptide anti- biotics surfactin, bacillomycin, fengycin and locillomycin were prepared by acid precipitation, organic solvent extraction and solid phase ex- traction from the culture broth of different Bacillus subtilis 916 and its nmtants; The purities of the four lipopeptides were detected by HPLC- MS and Tricine-SDS-PAGE ; The antifungal, antibacterial and hemolytic activities of the four lipopeptide antibiotics were analyzed by Fusari- mn oxyspomm, Staphylococcus aureus and sheep red blood cells. [ Result] The results of HPLC-MS and Tricine-SDS-PAGE showed that all the four family lipopeptide antibiotics prepared by this study reached electrophoresis pure and chromatography pure. Investigation of the bio- logical functions of the four class lipopepfides showed that surfactin and bacillomycin L had strong haemolytic activities, but fengycin and locillomycin only had weak haemolytic activities, while bacillomycin L and fengycin showed strong antifungal activities, but surfactin and locillomycin showed antibacterial and antiviral activities. [ Conclusion] In general, the efficient methods for purification of the four lipopeptide antibiotics surfactin, bacillomycin L, fengycin and locillomycin from Bacillus subtilis was set up and their biological activities were elucidet- ed, which laid a solid foundation to further elucidate the structure function relationship, biosynthesis pathway and application of frese li- popeptide antibiotics.
作者
罗楚平
张婧
季冬淳
陈树桥
陈永兴
赵玉萍
李相前
LUO Chu-ping;ZHANG Jing;JI Dong-chun;CHEN Shu-qiao;CHEN Yong-xing;ZHAO Yu-ping;LI Xiang-qian(College of Life Science and Food Engineering,Huaiyin Institute of Technology,Jiangsu Huai＇an 223002,China;Commerial College,Huaiyin Institute of Technology,Jiangsu Huai＇an 223002,China;Nanjing Fishery Research Institute,Jiangsu Nanjing 210036,China)
出处
《西南农业学报》
CSCD
北大核心
2018年第11期2307-2314,共8页
Southwest China Journal of Agricultural Sciences
基金
江苏省农业科技自主创新资金[CX(17)2008]
