生物学杂志

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产蛋白质类抑菌物质乳酸菌的分离鉴定与抑菌特性研究

  

  1. 1. 淮阴工学院 生命科学与食品工程学院,淮安 223003; 2. 淮阴工学院 化学工程学院,淮安 223003;3. 江苏省生物质转化与过程集成工程实验室,淮安 223003
  • 出版日期:2018-06-18 发布日期:2018-06-18
  • 通讯作者: 刘飞,博士,讲师,研究方向为环境微生物,E-mail:hyitliu@163.com
  • 作者简介:任世英,博士,副教授,研究方向为功能微生物,E-mail:rsy1314@163.com
  • 基金资助:
    国家自然科学基金项目(51308245);江苏省淮安市科技计划项目(HAN2015026);江苏省生物质转化与过程集成工程实验室开放课题(JPELBCPL2013004和JPELBCPL2014006);淮阴工学院大学生创新训练计划项目(201716015YJ)

Isolation and identification of lactic acid bacterium producing proteinaceous substances with antibacterial property and its antibacterial characteristics

  1. 1. School of Life Science and Food Engineering; 2. Faculty of Chemical Engineering;3. Jiangsu Provincial Engineering Laboratory for Biomass Conversion and Process Integration,Huaiyin Institute of Technology, Huaian 223003, China
  • Online:2018-06-18 Published:2018-06-18

摘要: 取传统发酵泡菜汤,通过透明圈法筛选出一株兼性厌氧型乳酸菌,菌落周围能产生抑菌圈,将其标记为A5。菌体呈短杆状,为革兰氏阳性菌,具有运动性。淀粉水解实验呈阴性,明胶液化实验、甲基红实验、葡萄糖氧化发酵实验、纤维素分解实验呈阳性。排除酸抑制作用和过氧化氢抑制作用后,发酵上清液依然维持较高的抑菌活性,而将发酵上清液用蛋白酶处理后,抑菌活性明显降低,综上实验说明,该乳酸菌A5发酵上清液中存在一种蛋白质类的抑菌物质(细菌素)。将发酵上清液的pH进行调节,然后测定其抑菌活性,发现在pH 2时仍然保持较强的抑菌活性,表明A5所产的抑菌物质对酸有良好的耐受性。经16S rDNA分子生物学鉴定,该菌属于鼠李糖乳杆菌属(Lactobacillus rhamnosus),暂命名为Lactobacillus rhamnosus sp. A5。该类物质对革兰氏阳性菌和革兰氏阴性菌有较好的抑制作用,具有较广的抑菌谱,而且对酸有良好的耐受性,为其开发成为一种天然无毒的广谱抑菌剂提供可能。

关键词: 泡菜, 乳酸菌, 抑菌圈, 细菌素

Abstract: A facultative anaerobic lactic acid bacterium producing antibacterial substrates was isolated from the traditional fermentation pickle by clear-circle method and marked as A5. Its cell is short-rod, G+, motile. The results of physiological and biochemical research were obtained that amylolysis test was negative, while gelatin liquefaction, methyl red, glucose fermentation and cellulose decomposition tests were positive. By excluding the influence of acid inhibition and hydrogen peroxide, the supernatant still had better antibacterial activity. When the supernatant was treated with protease, its antibacterial activity largely decreased. All the results indicated strain A5 fermentation supernatant contained a type of proteinaceous antibacterial substance (bacteriocin). By adjusting the pH value of supernatant and measuring its antibacterial activity, the result showed supernatant had stronger antibacterial activity at pH 2, which meant these antibacterial substances had super tolerance to acid. Strain A5 has good inhibitory activity to gram-positive bacteria and gram-negative bacteria, meaning its broad-spectrum antibacterial activity, and higher tolerance to acid, which make it a potential application in bio-antibiotics field.

Key words: pickle, lactic acid bacterium(LAB), inhibitory circle, bacteriocin