食品科学 ›› 2018, Vol. 39 ›› Issue (24): 175-182.doi: 10.7506/spkx1002-6630-201824027

• 生物工程 • 上一篇    下一篇

老白干酒曲中酵母菌多样性分析

文淑婷1,李?艳1,2,*   

  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)
  • 收稿日期:2018-12-25 修回日期:2018-12-25 出版日期:2018-12-25 发布日期:2018-12-17

Yeast Diversity Analysis of Laobaigan Liquor Daqu

WEN Shuting1, LI Yan1,2,*   

  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)
  • Received:2018-12-25 Revised:2018-12-25 Online:2018-12-25 Published:2018-12-17

摘要: 采用微生物分离培养与分子鉴定相结合的方法,在对酒曲中的酵母菌进行分离、纯化和形态学鉴定基础上,进行菌株核糖体26S rDNA聚合酶链式反应-限制性片段长度多态性分析技术结合的分子鉴定。并采用非培养方法,直接提取酒曲中的总DNA进行聚合酶链式反应扩增、电泳和切胶与回收,通过聚合酶链式反应-变性梯度凝胶电泳(polymerase chain reaction-denaturing gradient gel electrophoresis,PCR-DGGE)法鉴定酵母菌。结果表明,采用培养方式分离到57?株6?种不同形态的酵母菌,经鉴定为5?种分子类型,分别为奥默毕赤酵母(Pichia kudriavzevii)、葡萄牙棒孢酵母(Clavispora lusitaniae)、异常威克汉姆酵母(Wickerhamomyces anomalus)、胶红酵母(Rhodotorula mucilaginosa)、阿氏丝孢酵母(Trichosporon asahii)。经PCR-DGGE法鉴定得到10 株酵母菌,分别为W. anomalus、Wickerhamomyces sp.、T. asahii、热带假丝酵母(Candida tropicalis)和Candida sp.各1 株,Hyphopichia burtonii 2?株,另有uncultured Saccharomycopsis 3?株。培养与非培养两种方式共鉴定得到10 株酵母菌,分别属于8?个属,包括P. kudriavzevii、C. lusitaniae、W. anomalus、R. mucilaginosa、T. asahii、uncultured Saccharomycopsis、H. burtonii、Wickerhamomyces sp.、C. tropicalis、Candida sp.,该研究结果可为进一步研究酒曲的发酵能力提供参考。

关键词: 老白干酒曲, 酵母菌, 多样性, 26S rDNA聚合酶链式反应-限制性片段长度多态性, 聚合酶链式反应-变性梯度凝胶电泳

Abstract: In this study, yeasts were isolated and purified from the traditional fermentation starter (Daqu) for Laobaigan liquor. The isolates were identified by morphology as well as polymerase chain reaction-restriction fragment length polymorphism?(PCR-RFLP) analysis of ribosomal 26S rDNA. Meanwhile, yeasts were identified by culture-independent method, namely polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Total DNA was extracted from Daqu and amplified by PCR. The PCR products?were fingerprinted by denaturing gradient gel electrophoresis (DGGE) and were excised and recovered from the gel. The results showed that a total of 57 strains of yeast assigned to 6 kinds of morphology were isolated by traditional culture method and were identified as Pichia kudriavzevii, Clavispora lusitaniae, Wickerhamomyces anomalus, Rhodotorula mucilaginosa, and Trichosporon asahii. A total of 10 strains of yeast were obtained by PCR-DGGE, including one strain of W. anomalus, one strain of Wickerhamomyces sp., one strain of T. asahii, one strain of Candida tropicalis, one strain of Candida sp. two strains of Hyphopichia burtonii and three strains of uncultured Saccharomycopsis. Ten yeast strains belonging to eight genera were identified by culture-dependent and culture-independent methods, including P. kudriavzevii, C. lusitaniae, W. anomalus, R. mucilaginosa, T. asahii, uncultured Saccharomycopsis, H. burtonii, Wickerhamomyces sp., C. tropicalis, and Candida sp.. The results from this study provide a basis for further study on the fermentation performance of Daqu .

Key words: Laobaigan liquor Daqu, yeast, diversity, 26S rDNA PCR-RFLP, PCR-DGGE

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