酒鬼酒窖池酒醅芽孢杆菌的系统发育分析和胞外酶学特征  

Phylogenetic Analysis and Exoenzyme Properties of Bacilli Isolated from the Fermented Grains in the Fermentation Pits in Jiugui Liquor Distillery

作　　者：杨玉婷[1] 黎有有 吴秋蕾 樊林 刘荷[1] 赵湖 贺建武[1] 刘祝祥[1] 陈义光[1] YANG Yuting;LI Youyou;WU Qiulei;FAN Lin;LIU He;ZHAO Hu;HE Jianwu;LIU Zhuxiang;CHEN Yiguang(College of Biology and Environmental Sciences,Jishou University,Jishou,Hunan 416000;Jiugui Liquor Co.Ltd.,Jishou,Hunan 416000,China)

机构地区：[1]吉首大学生物资源与环境科学学院,湖南吉首416000 [2]酒鬼酒股份有限公司,湖南吉首416000

出　　处：《酿酒科技》2018年第10期97-102,共6页Liquor-Making Science & Technology

摘　　要：从酒鬼酒窖池中采集成熟酒醅样品,用纯培养法进行芽孢杆菌分离纯化,采用16S rRNA基因序列分析法进行系统发育分析,采用水解法对其胞外酶学特征进行观察。采用NA和LA培养基分离纯化得到104株细菌菌株,根据形态特征和产芽孢特征,去除部分冗余,选择28株代表性芽孢杆菌用于进一步研究。系统发育分析结果表明,这28株芽孢杆菌具有较高的类群多样性和物种多样性,属于细菌域(Bacteria)厚壁菌门(Firmicutes)的2个科(Bacillaceae、Paenibacillaceae)的3个属(Bacillus、Brevibacillus、Virgibacillus),分为10个物种。芽孢杆菌属(Bacillus)占绝对优势,共26株(92.9%);其余2株JSM161009和JSM 166025分属于枝芽孢杆菌属(Virgibacillus)和短芽孢杆菌属(Brevibacillus)。胞外水解酶学特征观察结果表明,至少产生一种酶的菌株共27株(96.4%)。其中有20株(71.4%)淀粉水解酶阳性,19株(67.9%)明胶水解酶阳性,27株(96.4%)尿素水解酶阳性,21株(75.0%)对4种吐温(20、40、60、80)之一水解活性阳性。在28株受试菌株中,有8株(JSM 161009、163010、163017、166014、167011、167021、168017、168019)水解酶谱较广。这8个菌株均属于芽孢杆菌科(Bacillaceae),其中1株(JSM161009)属于枝芽孢杆菌属(Virgibacillus),其他7株属于芽孢杆菌属(Bacillus)。In this study,the cultivable bacilli were isolated by culture-dependent method from the fermented grains in the fermentation pits in Jiugui Liquor Distillery.The diversity of the bacilli was investigated by phylogenetic analysis based on 16S rRNA gene sequence comparisons.Furthermore,the exoenzyme activities were studied by using some conventional methods.According to their morphology and the endospore observation results,28 representative bacilli out of 104 isolated bacilli were selected for phylogenetic analysis and exoenzyme observation.The results of phylogenetic analysis showed that the 28 bacilli represented 10 species belonging to 3 genera(Bacillus,Brevibacillus,and Virgibacillus)of 2 families(Bacillaceae and Paenibacillaceae)in 1 phyla(Firmicutes).Among the 28 bacilli,the dominant genus was Bacillus(26 strains,92.9%),and the other 2 strains,JSM 161009 and JSM 166025,belonged to Virgibacillus and Brevibacillus respectively.The exoenzyme observation showed that,27 out of 28(96.4%)bacilli pro-duced at least one enzyme,among them,20 strains(71.4%)were amylase-positive,19 strains(67.9%)were gelatinase-positive,27 strains(96.4%)were urease-positive,21 strains(75.0%)were esterase-positive(Tweens 20,40,60 and 80),and 8 bacilli strains(JSM 161009,163010,163017,166014,167011,167021,168017,168019)showed a relatively broad spectrum of exoenzymes.These 8 bacilli belonged to the family of Bacillaceae(strain JSM 161009 belonged to the genus Virgibacillus,and the other 7 strains belonged to the genus Bacillus).

关 键 词：酒鬼酒 酒醅 芽孢杆菌 系统发育分析 胞外酶 

分 类 号：TS262.3[轻工技术与工程—发酵工程] TS261.4[轻工技术与工程—食品科学与工程]