观音土曲乳酸菌的分离与分子鉴定  

Isolation and Molecular Identification of Lactobacillus from White Clay Distillers Yeast

作　　者：李睿[1] 沈汪洋[2] 翟平平[1] 陈道玉 

机构地区：[1]武汉工业学院生物与制药工程学院,湖北武汉430023 [2]武汉工业学院食品科学与工程学院,湖北武汉430023 [3]湖北劲牌有限公司,湖北大冶435100

出　　处：《安徽农业科学》2011年第4期1891-1892,1894,共3页Journal of Anhui Agricultural Sciences

基　　金：湖北省自然科学基金重点项目(2009CDA118)

摘　　要：[目的]指导白酒生产,提高白酒质量。[方法]采用平板分离法从观音土曲中分离到7株乳酸菌（B1~B7）,测定其发酵产乳酸的量。选取乳酸产量最高的1株菌B7,提取其DNA并进行16S rDNA的PCR扩增、克隆、测序。从GenBank中选取与B7同源性最高的典型菌株,下载其16S rDNA序列,采用邻位相连法构建进化树。[结果]B1~B7的产乳酸量分别为：24.7、20.7、30.7、15.2、9.5、21.8、35.7mg/100m l;对B7的基因组DNA进行16S rDNA的PCR扩增,并对纯化后的扩增产物进行TA克隆,阳性克隆经琼脂糖凝胶电泳后在3000和1 600 bp处出现电泳条带,说明B7的16S rDNA PCR扩增产物已克隆到测序载体上。同源性分析结果表明,B7为乳杆菌属的短乳杆菌。[结论]该研究从观音土曲中分离到1株高产乳酸菌——短乳杆菌。[Objective]The purpose of this study was to direct the production of distillate spirit and enhance its quality.[Method] Seven strains of lactobacilli（B1~B7） were isolated from white clay distillers yeast by plate isolation method and their lactic acid yields in fermentation was determined.The strain B7 with highest yield of lactic acid was selected for extracting DNA and performing PCR amplification,clone and sequencing on its 16S rDNA.The typical strains,which had highest homology with B7,were screened out from Genbank,their 16S rDNA sequences were loaded and the cladogram was constructed by ortho-linkage method.[Result] The lactic acid yields of B1~B7 were 24.7,20.7,30.7,15.2,9.5,21.8 and 35.7 mg/100ml.PCR amplification was performed on the 16S rDNA in genome DNA of B7 and TA clone was performed on purified amplification products.The electrophoresis strips of positive clones appeared at 3000 and 1600 bp in agarose gel electrophoresis,indicating that the PCR amplification products of 16S rDNA in B7 were cloned into sequencing vector.The results from homological analysis showed that B7 was Lactobacillus brevis of Acidobacterium.[Conclusion] Lactobacillus brevis-a strain of high-yield lactobacillus was isolated from white clay distillers yeast through this study.

关 键 词：观音土曲 16S RDNA 乳酸菌 分离 鉴定 

分 类 号：Q939.9[生物学—微生物学]