酒鬼酒制曲车间产淀粉酶细菌筛选和系统发育

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16SrRNA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16SrRNA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16SrRNA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.

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

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, Deinoeoccaceae, Micrococeaceae） 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 （Deinococ- cus-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.