传统分离培养结合PCR-DGGE技术分析传统乳制品中的乳酸菌  

Analysis of lactic acid bacteria in traditional dairy products by culture-independent and denaturing gradient gel electrophoresis(DGGE) methods

作　　者：董晓婉[1] 李宝坤[1] 卢士玲[1] 陈清敏[1] 鲁有国 常瀚泽 

机构地区：[1]石河子大学食品学院,新疆石河子832003

出　　处：《食品与发酵工业》2014年第3期97-101,共5页Food and Fermentation Industries

基　　金：新疆蒙古族传统发酵乳制品非发酵剂乳酸菌多样性及群落结构的研究;项目编号:31201395;新疆阿勒泰地区哈萨克族自然发酵乳制品乳酸菌种质资源评价及分子多态性研究(No.30960001);石河子大学高层次人才启动项目-传统发酵乳制品中增香型益生菌筛选及其功能(No.RCZX201223)

摘　　要：采用传统培养分离方法结合聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术对传统乳制品中乳酸菌种群结构进行研究。结果表明:用传统分离与分子鉴定方法得到5种乳酸菌,其中,乳酸片球菌(Pediococcus acidilactici)为优势菌群,对通过PCR-DGGE方法得到的9条16S rDNA条带序列进行了比对,结果表明瑞士乳杆菌(Lactobacillus helveticus)的丰度最高,是酸奶样品中主要的优势菌群,乳酸片球菌(Pediococcus acidilactici)为次优势菌群。传统分离法与PCR-DGGE技术结合能够更有效、更全面地分析传统乳制品中微生物的群落结构及优势菌群。In order to analyze lactic acid bacteria community structure in traditional dairy products and explore the predominant microbial species, both culture-dependent and culture-independent methods and denaturing gradient gel electrophoresis （DGGE） based on the sequence of 16S rDNA V6-V8 region gene were used to examine the total lactic acid bacteria counts. The results showed that 5 kinds of lactic acid bacteria were identified from traditional dairy products by the Culture-independent method, wherein the dominant bacteria belonged to Pediococcus acidilactici. By 16S rDNA V6-V8 region gene DGGE method, the most dominant bacteria was Lactobacillus helveticus, the second is Pediococcus acidilactici. This suggested that Independent culture combined with PCR-DGGE could be applied to analyze bacterial community structure and predominant bacteria in traditional dairy products effectively, thus provide a theoretical reference for the development of culture starter and that will give us more information of microorganism diversity.

关 键 词：乳酸菌 分离鉴定 16S RDNA 聚合酶链式反应-变性梯度凝胶电泳(PCR—DGGE) 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]