传统分离培养结合PCR-DGGE技术分析广式腊肠中优势菌  

Analysis of Dominant Microbial Species in Cantonese Sausage by Independent Culture and PCR-DGGE Technology

作　　者：谢科[1] 余晓峰 郑海松 宗凯 连英琪[1] 刘国庆[1] 

机构地区：[1]合肥工业大学生物与食品工程学院,安徽合肥230009 [2]安徽出入境检疫检验局,安徽合肥230022

出　　处：《食品科学》2013年第4期157-160,共4页Food Science

基　　金：国家质检总局科技计划项目(2010K135);安徽省战略型新兴产业重点科技攻关项目(11010402142)

摘　　要：采用传统培养方法结合聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)指纹技术对广式腊肠中微生物种群结构进行研究,分析广式腊肠发酵过程的优势微生物群落。结果表明:在传统分离培养中共得到葡萄球菌19株,乳酸菌12株,通过生理生化鉴定得到4株葡萄球菌(P1～P4)和2株乳酸菌(L1、L2)。DGGE指纹图谱显示广式腊肠混合菌群中有3条条带无对应的纯菌株而纯菌株中有1株在混合菌群的图谱上没有相应的条带。进一步的序列分析和相似性比较得出,P1、P4为木糖葡萄球菌,P2、P3、L1、L2分别为孔氏葡萄球菌、阿尔莱特葡萄球菌、格氏乳球菌、乳杆菌属。确定广式腊肠细菌菌群方面的主要优势菌为腐生葡萄球菌、乳杆菌属、木糖葡萄球菌。传统分离法与PCR-DGGE技术结合能够更有效、更全面地分析广式腊肠中微生物群落结构及优势菌。In order to analyze bacterial community structure in Cantonese sausages and explore the predominant microbial species, traditional culture combined with PCR-DGGE fingerprint technology was applied to investigate bacterial population structure in Cantonese sausages. The results showed that 19 Staphylococcus strains and 12 lactic acid strains were obtained by traditional culture method. Four Staphylococcus strains （named as P1-P4） and two lactic acid strains （named as L1 and L2） were identified by physiological-biochemical characteristics. Three bands of total bacterial DNA in DGGE profile of Cantonese sausages were not recovered by cultivation. Conversely, one bacterial species did not have the band of total bacterial DNA in DGGE fingerprint. Sequence analysis and similarity comparison showed that P1 and P4 were Staphylococcus xylosus, and P2, P3, L1 and L2 were Staphylococcus cohnii, Staphylococcus epidermidis, Lactococcus garvieae and Lactobacillales bacterium. The major predominant bacteria in Cantonese sausages were Staphylococcus saprophyticus, Lactobacillales bacterium and Staphylococcus xylosus. Independent culture combined with PCR-DGGE can be applied to the analysis of bacterial community structure and predominant bacteria in Cantonese sausages effectively, which will provide a theoretical reference for the development of culture starter.

关 键 词：广式腊肠 聚合酶链式反应-变性梯度凝胶电泳 16SrDNA 传统分离培养 优势菌 

分 类 号：Q503[生物学—生物化学]