食品与发酵工业 >

2022 , Vol. 48 >Issue 6: 270 - 275

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.027843

分析与检测

采用实时荧光定量PCR法检测白酒酿造系统中的重要功能菌株Lactobacillus jinshani

  • 张媛 ,
  • 廖卫芳 ,
  • 缪礼鸿 ,
  • 杨团元 ,
  • 刘蒲临 ,
  • 杨一斌
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  • 1(武汉轻工大学 生命科学与技术学院,湖北 武汉,430023)
    2(湖北白云边酒业股份有限公司,湖北 松滋,434200)
硕士(缪礼鸿教授为通信作者,E-mail:miaowhpu@126.com)

收稿日期: 2021-04-29

  修回日期: 2021-06-16

  网络出版日期: 2022-04-25

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Detection of important functional strain Lactobacillus jinshani in Baijiu brewing system by real-time fluorescence quantitative PCR

  • ZHANG Yuan ,
  • LIAO Weifang ,
  • MIAO Lihong ,
  • YANG Tuanyuan ,
  • LIU Pulin ,
  • YANG Yibin
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  • 1(College of Life Science and Technology, Wuhan Polytechnic University, Wuhan 430023, China)
    2(Baiyunbian Distillery Co.Ltd., Songzi 434200, China)

Received date: 2021-04-29

  Revised date: 2021-06-16

  Online published: 2022-04-25

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摘要

金山乳杆菌(Lactobacillus jinshani)是1株广泛存在于白酒酿造体系中的功能菌株,为了快速、准确地对白酒酿造过程中的L.jinshani进行定性、定量检测,通过设计L.jinshani特异性引物,建立了实时荧光定量 PCR 检测方法。该研究根据白酒酿造菌株L.jinshani 的全基因组序列设计了1对扩增产物大小为 266 bp 的特异性引物,经特异性、敏感性和重复性试验显示,在常见的13种白酒酿造微生物中,可特异性检测到L.jinshani,最低检出量为3.08×103 copies/μL。重复性结果变异系数均≤3.06%。进一步对白酒酿造中大曲及不同轮次酒醅样品进行了检测,结果显示,大曲及第1轮酒醅中未检测出L.jinshani,在第2轮次酒醅中L.jinshani含量达到(8.9±0.11) lgcopies/g,4轮酒醅中检出L.jinshani(9.38±0.11)lgcopies/g,第5轮酒醅中L.jinshani含量达到(9.43±0.11) lgcopies/g,与高通量测序所得结果一致。说明该实时荧光定量 PCR方法可实现对白酒酿造系统中L.jinshani的快速鉴定和定量检测。

关键词： 白酒; 实时荧光定量PCR; 乳酸菌; 金山乳杆菌; 定性检测; 定量检测

本文引用格式

张媛 , 廖卫芳 , 缪礼鸿 , 杨团元 , 刘蒲临 , 杨一斌 . 采用实时荧光定量PCR法检测白酒酿造系统中的重要功能菌株Lactobacillus jinshani[J]. 食品与发酵工业, 2022 , 48(6) : 270 -275 . DOI: 10.13995/j.cnki.11-1802/ts.027843

Abstract

Lactobacillus jinshani is a functional strain widely existing in the Baijiu brewing system. In order to rapidly and accurately detect the quantity and the quality of L. jinshani in Baijiu brewing, a quantitative real-time PCR (q-PCR) detection method was established by selecting and optimizing specific amplification primers of L. jinshani. In this study, a pair of specific primers with the size of 266 bp were designed according to the whole genome sequence of Baijiu-making strain L. jinshani. The tests of specificity, sensitivity and repeatability were carried out. Only L. jinshani could be detected among 13 common Baijiu brewing microorganisms, and the minimum detection amount was 3.08×103 copies/μL, with the coefficient of variation of reproducible results≤3.06%. Furthermore, the samples of Daqu and different rounds of fermented grains were tested, and it was found that no L. jinshani was detected in Daqu and the first round of fermented grains, the content of L. jinshani in the second round of fermented grains was (8.9±0.11) lgcopies/g, the content of L. jinshani in the fourth round of fermented grains was (9.38±0.11) lgcopies/g, and the content of L. jinshani in the fifth round of fermented grains reached (9.43±0.11) lgcopies/g, which was consistent with the results of high-throughput sequencing. The results showed that the fluorescence quantitative PCR method can be used for rapid identification and quantitative detection of L. jinshani in Baijiu brewing system.

Key words： Baijiu; q-PCR; Lactobacillus; Lactobacillus jinshani; qualitative detection; quantitative detection

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