源自泡菜的植物乳杆菌产新型广谱抑菌细菌素的特性分析

摘要/Abstract

摘要： 筛选对食源性致病菌和腐败菌有特异性抑制作用的产细菌素乳酸菌菌株，并研究细菌素的特性，为其在食品防腐中的应用奠定基础。从泡菜中分离的112株乳酸菌中反复筛选得到一株产细菌素（细菌素命名为BacH32）菌株（Hlh32），经鉴定菌株Hlh32为植物乳杆菌（Lactobacillus plantarum），其在35 ℃ MRS培养基中发酵至稳定期即18～24 h达到最大BacH32产量（对金黄色葡萄球菌的抗菌活力达到1850 Au/mL）。通过硫酸铵盐析、DEAE Sepharose Fast Flow离子交换层析、Sephadex G-15凝胶过滤层析、制备型高效液相色谱等方法进行纯化。Trinice SDS-PAGE结果显示BacH32分子质量为6.7 kDa，与文献报道的植物乳杆菌细菌素的分子量有显著差异。细菌素BacH32对蛋白酶敏感，蛋白酶K、木瓜蛋白酶、胰蛋白酶、胃蛋白酶均可使其抑菌活性完全丧失，而脂肪酶、α-淀粉酶和β-淀粉酶对其无影响，表明BacH32为蛋白质。BacH32在pH 2~9的环境下孵育4 h，在37 ℃、60 ℃、80 ℃、100 ℃保持30 min或121 ℃保持15 min仍能保持较好抑菌活性。细菌素BacH32对革兰氏阳性菌和革兰氏阴性菌具有广谱抑菌活性，如大肠杆菌、伤寒沙门氏菌、金黄色葡萄球菌、藤黄微球菌、粪肠球菌、绿脓杆菌、副溶血性弧菌、蜡样芽胞杆菌、枯草芽孢杆菌、单增李斯特氏菌等。此外，BacH32对假丝酵母、酿酒酵母、白色念珠菌等真菌也有一定的抑菌活性。向培养至对数初期的金黄色葡萄球菌中添加BacH32，菌体生长完全受到抑制。以上特性表明植物乳杆菌Hlh32合成的细菌素BacH32有潜力作为天然食品防腐剂在食品中使用。

关键词: 植物乳杆菌, 细菌素, 纯化, 抑菌作用

Abstract: To screen Lactobacillus strains which can produce specific inhibition to food borne pathogens and spoilage bacteria, and to further study the characteristics of bacteriocin, so as to lay a foundation for its application in food antisepsis. One of 112 isolates (Hlh32) produced a bacteriocin, designated as BacH32, and was isolated from Pickle juice and identified as Lactobacillus plantarum. This bacteriocin reached the maximum production （1850Au/mL） at phase stationary phase after 18-24 h of growth in MRS broth at 35 ℃ against Staphylococcus aureus. It was purified sequentially by performing ammonium sulfate precipitation, DEAE Sepharose Fast Flow anion exchange chromatography, Sephadex G-15 gel filtration chromatography and reversed-phase HPLC separation. According to Trinice SDS-PAGE, the molecular mass of bacteriocin Hlh32 was 6.7 kDa, which was significantly different from previous reported bacteriocins produced by Lactobacillus plantarum strains. It was sensitive to proteolytic enzymes. Complete inactivation of antimicrobial activity was observed after treatment of crude extracts with proteinase K, papain, trypsin, and pepsin, and the bacteriocin was resistant to treatments with lipase, α-amylase and β-amylase, which confirmed its proteinaceous nature. BacH32 remained active after 4 h at pH 2-9 and temperature treatments after 30 min at 37, 60, 80, 100 ℃, and 15 min at 121 ℃. Bacteriocin was found to have extensive spectrum antimicrobial activity against both Gram-positive and Gram-negative bacteria, such as Escherichia coli、Salmonella typhi、Staphylococcus aureus、Micrococcus luteus、Enterococcus faecalis、Pseudomonas aeruginosa、Vibrio parahaemolyticus、Bacillus cereus、Bacillus subtilis、Saccharomyces cerevisiae、Listeria monocytogenes. In addition, Hlh32 exhibited antimicrobial activity against fungus such as Candida krusei、Saccharomyces cerevisiae and Candida albicans. Addition of BacH32 to a mid-log culture of Staphylococcus aureus inhibited growth. These results indicate the potential of the bacteriocin for use as a food bio-preservative.

Key words: Lactobacillus plantarum, Bacteriocin, Purification, Antimicrobial characteristic

中图分类号:

Q819

高兆建黄亮浩丁飞鸿陈腾赵宜峰焦魏. 源自泡菜的植物乳杆菌产新型广谱抑菌细菌素的特性分析[J]. 食品科学, 0, (): 0-0.

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