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

doi:10.3969/j.issn.1007-2985.2014.05.012

Abstract

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

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.

References

[1] 陕小虎,敖宗华,沈才洪,等.中国固态白酒中酿酒微生物研究进展[C]//第12届中国科学技术协会年会(第3卷）.福州：中国科学技术协会年会，2010:1-6.

[2] 张双民.土壤中淀粉酶高产菌株的分离及产酶条件的优化[J].土壤肥料,2006(2):59-61.

[3] 赵湖.湘泉酒微生物群系的分布探讨[J].酿酒,2002,29(4):23-24.

[4] 陈义光,李文均,崔晓龙,等.具抗肿瘤活性放线菌菌株YIM 90022的分离和系统发育分析[J].微生物学报,2006,46(5):696-701.

[5] 于玺华,车凤翔.现代空气微生物学及采检鉴技术[M].北京：军事医学科学出版社,1998.

[6] 东秀珠,蔡妙英.常见细菌鉴定手册[M].北京:科学出版社.2001:43-65.

[7] HEMN-GMEZ S,ESPINOSA J C,UBEDA J F.Characterization of Wine Yeasts by Temperature Gradient Gel Electrophoresis (TGGE)[J].Issue FEMS Microbiology Letters,2000,193(1):45-50.

[8] CHUN J,LEE J,JUNG Y,WAGNER M.A Web-Based Tool for the Identification of Prokaryotes Based on 16S Ribosomal RNA Gene Sequences[J].International Journal of Systematic and Evolutionary Microbiology,2007,57(10):2 259-2 261.

[9] THOMPSON J D,GIBSON T J,PLEWNIAK F,et al.The CLUSTAL_X Windows Interface:Flexible Strategies for Multiple Sequence Alignment Aided by Quality Analysis Tools[J].Nucleic Acids Research,1997,25(24):4 876-4 882.

[10] KIMURA M.A Simple Method for Estimating Evolutionary Rates of Base Substitutions Through Comparative Studies of Nucleotide Sequences[J].Journal of Molecular Evolution,1980,16(2):111-120.

[11] TAMURA K,DUDLEY J,NEI M,et al.MEGA4:Molecular Evolutionary Genetics Analysis (MEGA) Software Version 4.0[J].Molecular Biology and Evolution,2007,24(8):1 596-1 599.

[12] FELSENSTEIN J.Confidence Limits on Phylogenies:An Approach Using the Bootstrap[J].Evolution,1985,39(4):783-791.

[13] 翁庆北,赵维娜,万晴姣,等.茅台酒曲中产淀粉酶细菌分离及性质[J].贵州师范大学学报:自然科学版,2005,23(2):20-28.

[14] GOYAL N,GUPTA J K,SONI S K.A Novel Raw Starch Digesting Thermostable α-Amylase from Bacillus Sp.Ⅰ-3 and Its Use in the Direct Hydrolysis of Raw Potato Starch[J].Enzyme Microb. Tech.,2005,37(7):723-734.

[15] SHIOU R J,HUNG H C,JEANG C L.Improving the Themostability of Raw Starch Digesting Amylase from a Cytophaga Sp. by Site Directed Mutagenesis[J].Appl. Environ. Microb.,2003,69(4):2 383-2 385.

[16] JEANG C L,CHEN LS,SHIOU R J,et al.Cloning of a Gene Encoding Raw Starch Digesting Amylase from Cytophaga Sp.and Its Expression in E. Coli[J].Appl. Environ. Microb.,2002,68(7):3 651-3 654.