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| 贝莱斯芽孢杆菌(Bacillus velezensis) MCCC 1A15695抑菌蛋白分离纯化及应用研究 |
| 付欢,李亚婷,肖贵谦,彭磊磊,夏金梅,邵宗泽,曹军伟,赖其良 |
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| (上海海洋大学海洋科学学院，上海 201306;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) |
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| 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。 |
| 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 |
| DOI：10.3969/J.ISSN.2095-4972.20230531002 |
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| 基金项目:科技部2021年科技基础资源调查专项(2021FY100903) |
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| Isolation and purification of bacteriostatic protein from Bacillus velezensisMCCC 1A15695 and its application |
| 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) |
| Abstract: |
| 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. |
| Key words: marine biology Bacillus velezensis antimicrobial protein antibacterial spectrum Vibrio glass post larval disease |