新疆冰川冰缘冻土低温淀粉酶部菌株分子生物学鉴定及酶学特性研究

Molecular Biologic Identification of Some Bacteria Strains Producing Low Temperature Amylase Isolated and Enzymatic Characterization from Frozen Soil in Xinjiang Glacier Ice Margin

【目的】对新疆冰川冰缘地区冻土中筛选出的具有良好产低温淀粉酶能力的菌株进行分子生物学鉴定,分析其酶学特性。【方法】用种子培养液活化菌株提取基因组DNA进行PCR扩增,将扩增成功菌株的PCR产物测序结果通过NCBI数据库比对,对冻土中产低温淀粉酶菌株的16S rDNA序列用MegAlign Fasta软件进行系统发育分析。利用Yoo改良法测低温淀粉酶酶活力。【结果】菌株A-2,A-3,A-4经16S rDNA序列分析获得的序列与数据库中解淀粉芽孢杆菌(Bacillus amyloliquefaciens strain NBRC 15535 16S ribosomal RNA,partial sequence(NR 04145))比对相似率达到99%。该菌株产生的低温淀粉酶的最适作用温度为30℃,最适pH为6。【结论】菌株A-2,A-3,A-4均属解淀粉芽孢杆菌。

Objective] In this paper,some bacterial strains producing low temperature amylase isolated from frozen soil in Xinjiang periglacial zone were identified by molecular biology method and analysis of its enzymatic characterization.[Method] Genome DNA was extracted from bacterial strains activated in seed nutrient solution,followed by PCR amplification and the product was compared by NCBI data base,the 16S rDNA of bacteria strains producing low temperature-amylase was analyzed by MegAlign Fasta,using improved Yoo method to measure low temperature amylase enzyme.[Result]The result showed that similarity rules of the sequences between A-2,A-3,A-4 bacterial strains analyzed by 16S rDNA and the Bacillus amyloliquefaciens database were up to 99%.This strain produced the optimum temperature amylase is 30℃,and the optimum pH is 6.[Conclusion] The conclusion was that bacteria strains of A-2,A-3,A-4 were all Bacillus amyloliquefaciens.