贝莱斯芽孢杆菌(Bacillus velezensis)MCCC 1A15695抑菌蛋白分离纯化及应用研究  

Isolation and purification of bacteriostatic protein from Bacillus velezensis MCCC 1A15695 and its application

作　　者：付欢 李亚婷 肖贵谦 彭磊磊 夏金梅 邵宗泽 曹军伟 赖其良 FU Huan;LI Yating;XIAO Guiqian;PENG Leilei;XIA Jinmei;SHAO Zongze;CAO Junwei;LAI Qiliang(College of Marine Sciences,Shanghai Ocean University,Shanghai 201306,China;Key Laboratory of Marine Genetic Resources,Third Institute of Oceanography,MNR,Xiamen 361005,China)

机构地区：[1]上海海洋大学海洋科学学院,上海201306 [2]自然资源部第三海洋研究所、自然资源部海洋生物遗传资源重点实验室,福建厦门361005

出　　处：《应用海洋学学报》2024年第2期217-224,共8页Journal of Applied Oceanography

基　　金：科技部2021年科技基础资源调查专项(2021FY100903)。

摘　　要：对虾“玻璃苗”病的暴发,会导致对虾幼苗大批死亡,给水产养殖带来巨大损失。利用本实验室最近鉴定的“玻璃苗”致病菌副溶血弧菌(Vibrio parahaemolyticus)vp-HL,从海洋来源芽孢杆菌中筛选拮抗菌,获得一株能够高效抑制vp-HL菌株的贝莱斯芽孢杆菌(Bacillus velezensis)MCCC 1A15695。经分离鉴定与活性测定,其有效抑菌物质为蛋白类,该蛋白抑菌谱较广,对包括多种弧菌病原在内的17种病原微生物有抑制作用。经Q Seharose Fast Flow阴离子柱层析、聚丙烯酰胺凝胶电泳(SDS-PAGE)及液相色谱-质谱联用技术(LC-MS)分离纯化鉴定,确定活性蛋白组分中的有效抑菌物质为枯草杆菌素(subtilisin)、未知蛋白(hypothetical protein)及α-淀粉酶(Alpha-amylase)。蛋白类物质具有高生物安全性,本研究为后续益生菌的开发利用提供了理论基础。The outbreak of shrimp“glass post-larval disease”has led to a large number of diseases of shrimp larvae,bringing huge losses to aquaculture.Using the recently identified Vibrio parahaemolyticus vp-HL of the“glass post-larval disease”pathogen Vibrio parahaemolyticus,the antagonistic antibacterial screening of Bacillus of marine origin was carried out to obtain a strain of Bacillus velezensis MCCC 1A15695 that can effectively inhibit the strains MCCC 1A15695.After isolation and activity determination,the effective bacteriostatic substances were proteins,with a wide inhibitory spectrum,and had inhibitory effects on 17 pathogenic microorganisms,including a variety of Vibrio pathogens.After Q Seharose Fast Flow anion column chromatography,polyacrylamide gel electrophoresis(SDS-PAGE)and liquid chromatography-mass spectrometry(LC-MS),the effective bacteriostatic substances in the active protein components were subtilisin,hypothetical protein and Alpha-amylase.In view of the high biosafety of proteins,the results provide a theoretical basis for the subsequent development and utilization of probiotics.

关 键 词：海洋生物学 贝莱斯芽孢杆菌 抗菌蛋白 抑菌谱 弧菌 玻璃苗 

分 类 号：P735[天文地球—海洋生物学]