筛选自泡菜的发酵乳杆菌细菌素纯化及抑菌特性分析

从中国传统发酵蔬菜中筛选鉴定对食源性致病菌有广谱抗性的细菌素（命名为BLF52）产生菌株并对细菌素分离纯化和抑菌特性分析。分子筛凝胶过滤层析和半制备高效液相色谱（RP-HPLC）纯化细菌素并用Tricine-SDS-PAGE法测定其分子量。筛选得到产细菌素的发酵乳杆菌（Lactobacilus fermentum） CH-16。MRS培养基中34℃发酵至稳定期前期即24~30 h对S.aureus抑制活性最大（875 AU/mL）。BLF52纯化至单一组分，分子量5.6 kDa，40~80℃（30 min）、pH2.0~7.0（2 h）有良好稳定性。胃蛋白酶、木瓜蛋白酶、胰蛋白酶和蛋白酶K处理后，抑菌活性完全丧失，α-淀粉酶对其无影响，表明BLF52有典型的细菌素特性。BLF52对食源性腐败菌和致病菌如单增李斯特菌、金黄色葡萄球菌、蜡状芽孢杆菌、鼠伤寒沙门氏菌、铜绿假单胞菌、大肠杆菌和副溶血性弧菌均有抑制活性，通过杀菌模式抑制培养中的S.aureus生长。细菌素BLF52有潜力开发为适合酸性或中性食品使用的食品防腐剂。

Purification and Antibacterial Properties of Bacteriocin Produced by Lactobacilus fermentum Isolated from Pickled Vegetables

The aim of this study was to screen and identify strains producing bacteriocin(named BLF52)with broad-spectrum resistance to foodborne pathogens from Chinese traditional fermented vegetables,and to isolate and purify the bacteriocin and analyze its antibacterial characteristics.Molecular sieve gel filtration chromatography and semi-preparative high-performance liquid chromatography(RP-HPLC)were used to purify bacteriocin and the molecular weight was determined by Tricine-SDS-PAGE.A bacteriocin-producing Lactobacilus fermentum CH-16 strain was isolated.The highest inhibition of S.aureus(875 AU/mL)was noted at early stationary phase after 24~30 h of growth in MRS medium at 34℃.BLF52 was purified to homogeneity and its molecular mass was 5.6 kDa.It was stable in the range of 40~80℃(30 min)and pH2.0~7.0(2 h).After treatment with pepsin,papain,trypsin and proteinase K,the antibacterial activity of BLF52 was completely lost,andα-amylase had no effect on it,indicating that BLF52 had typical bacteriocin characteristics.BLF52 exhibited a broad antimicrobial spectrum against food-borne spoilage and pathogenic bacteria such as Listeria monocytogenes,Staphylococcus aureus,Bacillus cereus,Salmonella typhi,Pseudomonas aeruginosa,Escherichia coli,and Vibrio parahaemolyticus.Bacteriocins BLF52 inhibitted the growth of S.aureus in culture through bactericidal mode.BLF52 has the potential to be developed as a food preservative suitable for acidic or neutral foods.