食品科学

• 生物工程 • 上一篇    下一篇

羊羔美酒大曲中乳酸菌多样性及分子鉴定

李 艳1,2,董振玲1,3,李 佳1,牟德华1,*   

  1. 1.河北科技大学生物科学与工程学院,河北 石家庄 050018;2.河北省发酵工程技术研究中心,河北 石家庄 050018;3.河北一然生物科技有限公司,河北 石家庄 050800
  • 出版日期:2015-07-15 发布日期:2015-07-08

Diversity and Molecular Biological Identification of Lactic Acid Bacteria from Yanggaomeijiu Daqu, a Traditional Chinese Liquor Fermentation Starter

LI Yan1,2, DONG Zhenling1,3, LI Jia1, MOU Dehua1,*   

  1. 1. College of Bioscience and Bioengineering, Hebei University of Science and Technology, Shijiazhuang 050018, China;
    2. R&D Center for Fermentation Engineering of Hebei Province, Shijiazhuang 050018, China;
    3. Hebei Inatural Biological Technology Co. Ltd., Shijiazhuang 050800, China
  • Online:2015-07-15 Published:2015-07-08

摘要:

目的:分离和鉴定羊羔美酒大曲中的乳酸菌,探寻其菌群多样性,为酿酒工艺条件改进提供参考。方法:将酒曲进行梯度稀释、富集培养、平板画线等分离纯化,获取乳酸菌单菌落。采用菌落特征和个体形态特征结合的方法进行乳酸菌形态鉴定。乳酸菌的分子鉴定采用重复序列聚合酶链式反应(repetitive sequence-basedpolymerase chain reaction,Rep-PCR)技术和16S rDNA序列分析法。结果:从羊羔美酒大曲中共得到65 株乳酸菌,形态学分为9 类。用Rep-PCR技术在75%的相似性上将其区分为5 类,经基因序列分析,鉴定为分属于4 个属的乳酸菌,分别为:片球菌属(Pediococcus)、明串珠菌属(Leuconostoc)、乳杆菌属(Lactobacillus)、肠球菌属(Enterococcus)。结论:传统微生物分离技术与现代分子鉴定技术相结合,可快速准确鉴定出羊羔美酒大曲中乳酸菌的菌群组成和多样性。

关键词: 羊羔美酒大曲, 乳酸菌, 分子鉴定, 重复序列聚合酶链式反应, 16S rDNA序列分析

Abstract:

Purpose: The aim of this work was to isolate and identify lactic acid bacteria (LAB) in Yanggaomeijiu Daqu,
a fermentation starter for the production of Chinese liquor Yanggaomeijiu, and to explore the diversity of LAB in
Yanggaomeijiu Daqu. Methods: Individual bacterial colonies of LAB from Yanggaomeijiu Daqu were obtained through
isolation and purification steps including gradient dilution, enrichment, cultivation, and plate streaking. Morphological identification
of LAB isolates was performed based on colony and individual morphological characteristics and further molecular identification
was carried out by repetitive sequence-based polymerase chain reaction (Rep-PCR) and 16S rDNA sequence analysis. Results: A
total of 65 LAB isolates were obtained from Yanggaomeijiu Daqu, which could be assigned into 9 different groups by conventional
microbiological analysis, differentiated into five types by Rep-PCR at a 75% similarity, and belonged to 4 genera including
Pediococcus,Leuconostoc, Lactobacillus and Enterococcus by gene sequencing. Conclusion: The diversity and composition
of LAB in Yanggaomeijiu Daqu can be identified quickly and accurately by the combination of traditional microbial separation
methods and modern molecular biological identification techniques.

Key words: Yanggaomeijiu, lactic acid bacteria, molecular biological identification, repetitive sequence-based polymerase chain reaction (Rep-PCR), 16S rDNA sequence analysis

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