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(Clavisporalusitaniae)、异常威克汉姆酵母(Wickerhamomycesanomalus)、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,Hyphopichiaburtonii 2株,另有unculturedSaccharomycopsis3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。
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 ribosomal26S 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 of57strains of yeast assigned to6kinds 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 of10strains 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.
作者
文淑婷
李艳
WEN Shuting;LI Yan(College of Bioscience and Bioengineering, Hebei University of Science and Technology, Shijiazhuang 050018, China;R&D Center for Fermentation Engineering of Hebei Province, Shijiazhuang 050018, China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2018年第24期175-182,共8页
Food Science
