酒鬼酒制曲车间产淀粉酶细菌筛选和系统发育

doi:10.3969/j.issn.1007-2985.2014.05.012

journal6 ›› 2014, Vol. 35 ›› Issue (5): 51-54.DOI: 10.3969/j.issn.1007-2985.2014.05.012

酒鬼酒制曲车间产淀粉酶细菌筛选和系统发育

钟小娟,余冰,刘荷,肖建青,李洪军,樊林,程金莲,刘祝祥,陈义光

(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)

出版日期:2014-09-25 发布日期:2014-10-30
通讯作者: 陈义光（1965-），男，湖南新化人，吉首大学生物资源与环境科学学院教授，硕士生导师，主要从事微生物资源与分类研究.
作者简介:钟小娟（1983-），女（白族），湖南桑植人，吉首大学生物资源与环境科学学院硕士研究生，主要从事微生物资源与生态研究
基金资助:
国家自然科学基金资助项目（30970007）；湖南省重点学科建设资助项目（JSU071312Z01）；湖南省高校科技创新团队支持计划资助项目（201208Z01）；酒鬼酒股份有限公司委托项目（JGKJ 2012-JD03）；吉首大学校级科研课题资助项目（13JDY017）

Screening and Phylogenetic Analysis of Amylase-Producing Strains Isolated from the Air Samples from a Liquor Starter-Making Workshop of Jiugui Liquor Co. Ltd.

ZHONG Xiao-Juan, YU Bing, LIU He, XIAO Jian-Qing, LI Hong-Jun, FAN Lin, CHENG Jin-Lian, LIU Zhu-Xiang, CHEN Yi-Guang

(1.College of Biology and Environmental Sciences,Jishou University,Jishou 416000,China;2.Jiugui Liquor Co. Ltd.,Jishou 416000,China)

Online:2014-09-25 Published:2014-10-30

摘要/Abstract

摘要：采用平板水解圈法对酒鬼酒制曲车间空气的细菌进行产淀粉酶活性筛选，运用基于16S rRNA基因序列的分析法对高酶活菌株进行系统发育分析.研究结果表明，65株受试菌株中，有24.6%的菌株（16株）淀粉水解酶活性呈阳性，其中有4株(JSM 2145008，JSM 2155001，JSM 2155017，JSM 2175012)产淀粉水解酶能力较强.基于16S rRNA基因序列的系统发育分析表明，这4个菌株分别属于细菌域3个大的系统发育类群中3个科的3个属.菌株JSM 2145008属于放线菌门微球菌科，与节杆菌属（Arthrobacter）的氧化节杆菌的系统发育关系最为密切，它们之间的16S rRNA基因序列相似性高达99.8%，且在系统进化树上形成一个稳定的亚分支；菌株JSM 2155001和JSM 2175012属于异常球菌——栖热菌群异常球菌科的异常球菌属（Deinococcus），分别与该属的大琼异常球菌和云威异常球菌系统发育关系最为密切（序列相似性分别为99.6%和99.9%）；菌株JSM 2155017属于厚壁菌门芽孢杆菌科，与芽孢杆菌属（Bacillus）的同温层芽孢杆菌系统发育关系最为密切（序列相似性为100%），它们以极高的自展值支持（boostrap value,100%）在系统进化树上聚在一起.上述实验结果表明，酒鬼酒制曲车间空气源细菌中存在较高比例的产淀粉酶菌株，且这些菌株具有较高的类群多样性.

关键词： 酒鬼酒制曲车间, 空气源微生物, 淀粉酶活性, 系统发育分析

Abstract: The culturable bacteria in the air samples from a liquor starter-making workshop in Jiugui Liquor Co.Ltd.,China were isolated by using the natural precipitation method and culture-dependent method.The amylase-producing ability of the strains was investigated by using the plate hydrolysis spot method.The results showed that 16 out of 65 (24.6%) strains tested exhibited amylase-producing activity,in which 4 strains (JSM 2145008,JSM 2155001,JSM 2155017,JSM 2175012) could relatively strongly produce amylase.Subsequently,these 4 strains were investigated by means of a phylogenetic analysis based on 16S rRNA gene sequence comparisons.The results showed that these 4 strains were the members of 3 families (Bacillaceae,Deinococcaceae,Micrococcaceae) in 3 phylogenetic group/phyla (Actinobacteria,Deinococcus-Thermus,Firmicutes):strain JSM 2145008 belonged to the genus Arthrobacter (Actinobacteria,Micrococcaceae),being most closely to the type strain of Arthrobacter oxydans (sequence similarity 99.8%);strains JSM 2155001 and JSM 2175012 were the members of the genus Deinococcus (Deinococcus-Thermus,Deinococcaceae),being related most closely to the type strains of Deinococcus daejeonensis (99.6%) and Deinococcus yunweiensis (99.9%),respectively;strain JSM 2155017 belonged to the genus Bacillus (Firmicutes,Bacillaceae),exhibiting 100% 16S rRNA gene sequence similarity to the type strain of Bacillus stratosphericu.In conclusion,there is abundant bacterial diversity,as well as a number of amylase-producing strains in the air samples from a liquor starter-making workshop in Jiugui Liquor Co. Ltd.

Key words: liquor starter-making workshop, airborne microbes, amylase activity, phylogenetic analysis

钟小娟, 余冰, 刘荷, 肖建青, 李洪军, 樊林, 程金莲, 刘祝祥, 陈义光. 酒鬼酒制曲车间产淀粉酶细菌筛选和系统发育[J]. journal6, 2014, 35(5): 51-54.

ZHONG Xiao-Juan, YU Bing, LIU He, XIAO Jian-Qing, LI Hong-Jun, FAN Lin, CHENG Jin-Lian, LIU Zhu-Xiang, CHEN Yi-Guang. Screening and Phylogenetic Analysis of Amylase-Producing Strains Isolated from the Air Samples from a Liquor Starter-Making Workshop of Jiugui Liquor Co. Ltd.[J]. journal6, 2014, 35(5): 51-54.

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酒鬼酒制曲车间产淀粉酶细菌筛选和系统发育

Screening and Phylogenetic Analysis of Amylase-Producing Strains Isolated from the Air Samples from a Liquor Starter-Making Workshop of Jiugui Liquor Co. Ltd.

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