芽孢杆菌ZYCHH-01的鉴定与分泌产物纯化及其抑菌机理  

Identification,Purification of Secretory Product and Antibacterial Mechanism of Bacillus ZYCHH-01

作　　者：蔡巧贤 彭倩 李容博 肖婷 杨志东 高亚惠 赵逸凡 吴夏泠 张玉苍 CAI Qiaoxian;PENG Qian;LI Rongbo;XIAO Ting;YANG Zhidong;GAO Yahui;ZHAO Yifan;WU Xialing;ZHANG Yucang(College of Ocean Food and Biological Engineering,Jimei University,Xiamen 361021,China)

机构地区：[1]集美大学海洋食品与生物工程学院,福建厦门361021

出　　处：《集美大学学报（自然科学版）》2024年第4期308-320,共13页Journal of Jimei University：Natural Science

基　　金：国家自然科学基金面上项目“甲壳素常压催化液化机理及液化产品的应用研究”(21978059);科技部重点研发计划项目“多源性配料制备及营养设计”(2021YFD210020202)、“馅料类方便主食品质提升及新产品创制关键技术与示范”(2021YFD210020404);福建省中青年教师教育科研项目“探究改良果胶酶温度耐受性及其应用”(JAT200280);集美大学科研启动项目“蛋白质-壳聚糖复合水凝胶的设计和应用”(ZQ2021016)。

摘　　要：对实验室自主分离的芽孢杆菌ZYCHH-01进行多相分类和16S rRNA测序,确定其种属;通过乙酸乙酯萃取法、高效液相色谱纯化、高分辨四极杆飞行时间质谱联用、扫描电子显微镜和碱性磷酸酶泄露等方法,确定抑菌物质,探究其抑菌机理。实验结果表明,芽孢杆菌ZYCHH-01纯化产物可以抑制金黄色葡萄球菌、单核增生李斯特菌等革兰氏阳性菌,大肠杆菌、福氏志贺氏菌、鼠伤寒沙门氏菌、副溶血弧菌等革兰氏阴性菌,以及总状毛霉、黑根霉等真菌的生长,有效抑菌物质的相对分子质量为1072.6862,1058.6672,1044.6511,1030.6395 u的表面活性素同系物。此外,实验结果还表明,芽孢杆菌ZYCHH-01的抑菌机理可能是破坏细菌细胞壁。可见,芽孢杆菌ZYCHH-01是一株贝莱斯芽孢杆菌,有效抑菌物质是14~16个碳原子的表面活性素,能抑制多种革兰氏阳性菌、革兰氏阴性菌和真菌的生长。The traditional classification method and 16S rRNA gene sequencing were used to identify Bacillus ZYCHH-01 strain isolated in our laboratory.The crude bacteriocin extract was obtained by ethyl acetate extraction,and the antibacterial components were confirmed by high performance liquid chromatography and quadrupole time-of flight mass spectrometer.Based on the scanning electron microscope and alkaline phosphatase leakage experiment,the antibacterial mechanism of antibacterial substances was analyzed.The fermentation liquid and extraction of Bacillus ZYCHH-01 could inhibit the growth of gram-positive bacteria including Staphylococcus aureus and Listeria monocytogene s;Gram-negative bacteria such as Escherichia coli,Shigella flexneri,Salmonella typhimurium and Vibrio parahaemolyticus and fungi containing Mucor racemosus and Rhizopus nigricans.The mass spectral molecular weight of the effective antibacterial components were 1072.6862,1058.6672,1044.6511,1030.6395 u,respectively.The antibacterial substances produced by Bacillus ZYCHH-01 may inhibit bacterial growth by disturbing synthesis of bacterial cell wall.The Bacillus ZYCHH-01 was Bacillus velezensis,and the antibacterial components were surfactin containing 14-16 carbon atoms and its homologs.

关 键 词：芽孢杆菌 分泌物纯化 表面活性素 抑菌机理 多相分类 

分 类 号：Q939[生物学—微生物学]