食品科学 ›› 2021, Vol. 42 ›› Issue (10): 171-177.doi: 10.7506/spkx1002-6630-20200213-135

• 生物工程 • 上一篇    下一篇

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

高兆建,黄亮浩,丁飞鸿,赵宜峰,陈腾   

  1. (1.徐州工程学院食品与生物工程学院,江苏 徐州 221018;2.长江桂柳食品睢宁有限公司,江苏 徐州 221000)
  • 出版日期:2021-05-25 发布日期:2021-06-02
  • 基金资助:
    江苏省高等学校自然科学研究重大项目(20KJA180008); 江苏省苏北科技计划项目(XZ-SZ201819;BC2013417;BN2015021);徐州市科技计划项目(KC17083)

Characterization of Novel Broad-Spectrum Bacteriocin Produced by Lactobacillus plantarum from Pickles

GAO Zhaojian, HUANG Lianghao, DING Feihong, ZHAO Yifeng, CHEN Teng   

  1. (1. School of Food and Biological Engineering, Xuzhou University of Technology, Xuzhou 221018, China; 2. Yangtze River Guiliu Food Suining Co. Ltd., Xuzhou 221000, China)
  • Online:2021-05-25 Published:2021-06-02

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提供参考。从分离自泡菜的112 株乳酸菌中反复筛选得到1 株产细菌素(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抑菌活性达到1 850 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十二烷基硫酸钠-聚丙烯酰氨凝胶电泳结果显示BacH32分子质量为6.7 kDa,与文献报道的植物乳杆菌细菌素分子质量有显著差异。细菌素BacH32对蛋白酶敏感,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,对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: This research aimed to screen for Lactobacillus strains with the capability of producing bacteriocin that can specifically inhibit foodborne pathogens and spoilage bacteria and to study the characteristics of the bacteriocin, so as to lay a foundation for its application in food preservation. Out of 112 isolates from pickles, one, coded Hlh32, was found able to produce bacteriocin (BacH32) and identified as Lactobacillus plantarum. The maximum production of BacH32 (anti-Staphylococcus aureus activity of 1 850 AU/mL) was achieved when the culture reached the stationary phase (between 18 and 24 h) in MRS broth at 35 ℃. The bacteriocin was purified sequentially by ammonium sulfate salting-out, DEAE Sepharose Fast Flow anion exchange chromatography, Sephadex G-15 gel filtration chromatography and reversed-phase HPLC. As determined by Trinice SDS-PAGE, the molecular mass of bacteriocin Hlh32 was 6.7 kDa, which was significantly different from that previously reported. It was sensitive to proteolytic enzymes but to lipase, α-amylase not β-amylase, indicating its protein nature. Its antimicrobial activity was maintained well after incubation for 4 h at pH 2–9, 30 min at 37, 60, 80 and 100 ℃, or 15 min at 121 ℃. The bacteriocin was found to have board-spectrum antimicrobial activity against both Gram-positive and Gram-negative bacteria together with fungi. BacH32 completely inhibited the growth of S. aureus cells at the early-log phase. These results indicate the potential of the bacteriocin for use as a food biopreservative.

Key words: Lactobacillus plantarum; bacteriocin; purification; antimicrobial characteristics

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