枯草芽孢杆菌抑菌活性物质鉴定、抑菌特性及发酵条件优化  

Identification and Characterization of Antibacterial Metabolites and Optimization of Cultural Conditions for Bacillus subtilis

作　　者：张雯[1] 卞丹 沈燕秋 王文思 刘志彬[1] 倪莉[1] 

机构地区：[1]福州大学食品科学技术研究所,福州350108

出　　处：《中国食品学报》2017年第12期105-115,共11页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金-青年科学基金项目(31601532);福建省自然科学基金面上项目(2016J01156);福州市科技局院校合作项目(2016-G-75)

摘　　要：对具有抑菌活性的枯草芽孢杆菌BS08菌株进行抑菌活性物质的鉴定,抑菌特性研究和发酵条件优化。结果表明,枯草芽孢杆菌BS08发酵液经酸沉醇提的抑菌粗提物,再经薄层层析纯化、富集,红外吸收光谱测定为内酯类的环状脂肽,高效液相色谱和质谱鉴定其分子质量在995~1 350之间,有效成分为枯草芽孢杆菌表面活性素和伊枯草菌素。抑菌粗提物有较广的抑菌谱,对多种鱼源腐败菌有显著的抑制作用,该物质还具有良好的热稳定性和酸碱稳定性。在酵母浸膏14 g/L,无水葡萄糖14 g/L,硫酸镁1 g/L的营养液中,选择接种量7.0%,pH 8.2,培养温度38℃,200 r/min发酵24 h,结果发酵液中脂肽类物质含量达450 mg/L,抑菌率达69.6%。The active metabolites of Bacillus subtilis BS08 against bacterium purified from fish were identified by Infrared Spectroscopy and HPLC-MS. Liquid-state fermentation by BS08 was optimized for the maximum antibacterial activity by response surface methodology（RSM）. The active metabolites were extracted by acid and alcohol, purified by TLC and identified by Infrared Spectroscopy and HPLC-MS. The analysis revealed the active metabolites were families of lipopeptides, such as surfactin and iturin with molecular mass 995-1 350, thermal and pH stability, widely antimicrobial spectrum. Response surface methodology（RSM） was employed to optimize the cultural conditions of BS08 for the enhancement of lipopeptides. BS08 with 7% inoculum was cultivated at 38 ℃,200 r/min, pH 8.2 for 24 h in containing yeast extract 14 g/L, glucose 14 g/L, Mg SO41 g/L. The yield of lipopeptides reached 450 mg/L.

关 键 词：枯草芽孢杆菌BS08 发酵条件优化 脂肽 抑菌 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]