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动物乳杆菌的分离鉴定及其抑菌蛋白的特性分析

Screening and Identification of Lactobacillus Animalis Strain and Its Inhibitory Protein Characteristics

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【作者】 李明雄李妮李征向文良杨志荣罗璠

【Author】 LI Ming-Xiong1 LI Ni1 LI Zheng1 XIANG Wen-Liang1 YANG Zhi-Rong1 LUO Fan1,2* (1. Key Laboratory of Microbial-resource and Microbial-technique, College of Life Sciences, Sichuan University, Chengdu, Sichuan 610064, China) (2. College of Life Sciences and Technoligy, Southwest University for Nationalitise, Chengdu, Sichuan 610064, China)

【机构】 四川大学生命科学学院四川省资源微生物与微生物技术重点实验室西南民族大学生命科学与技术学院

【摘要】 通过滤纸片法从健康肉猪猪大肠、小肠中分离得到212株抗生物质产生菌,以杯碟法复筛,得到1株对溶壁微球菌(Micrococcus lysodeikticus)、金黄色葡萄球菌(Staphylococcus aureus)、枯草芽孢杆菌(Bacillus subtilis)、蜡状芽孢杆菌(Bacillus cereus)等革兰氏阳性菌和大肠杆菌(Escherichia coli)、鼠伤寒沙门氏菌(Salmonella typhimurium)等革兰氏阴性菌以及部分真菌如禾谷镰刀霉(Fusarium graminearum)均有强烈抑制作用的乳酸菌。经形态学、生理生化特征及16SrDNA序列同源性分析等手段鉴定该菌株为动物乳杆菌(Lactobacillus animalis)。排除酸和过氧化氢的干扰后,该菌株的发酵上清液对指示菌仍有明显抑菌活性;用蛋白酶处理该菌株的发酵上清液后,抑菌活性丧失;发酵液粗提物具有较好的热稳定性(经121°C处理20min仍有较强抑菌活性)以及较宽的抑菌活性pH值范围(3.5~5.5),因此初步认为该菌株产生一类具有广谱抑菌活性的类细菌素物质。

【Abstract】 A bacteriocin-like substance producing strain T12 was screened from 212 lactic acid bacteria isolated from intestine of healthy pigs by paper disk test and was identified as Lactobacillus. The supernatant of T12 exhibited strong inhibitory activity against the gram-positive bacteria, including Micrococcus lysodeikticus, Staphylococcus aureus, Bacillus subtilis and Bacillus cereus. A strong activity was also observed against the gramnegative bacteria and some eumycete, such as Escherichia coli, Salmonella typhimurium and Fusarium graminearum. Through detection of its appearance, physiological and biochemical characteristics and 16S rDNA gene sequence homology analysis, it was identified as Lactobacillus animalis. Eliminated the organic acids and hydrogen peroxide the fermented production exhibited strong inhibitory activityagainst indicat bacterias still; the inhibitory activity was totally lost after treatment with protease; The bacteriocin-like substance was heat-stable even at the autoclaving temperature (121°C for 20 min) and was active over a wide pH range (3.5~5.5). The protein nature and restricted spectrum of the substance points to its being bacteriocin-like substance.

【基金】 四川省科技攻关项目(No.07SG0832)
  • 【文献出处】 微生物学通报 ,Microbiology , 编辑部邮箱 ,2009年07期
  • 【分类号】Q93
  • 【被引频次】13
  • 【下载频次】433
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