巴斯德醋酸杆菌AC2005乙醇脱氢酶基因克隆与蛋白序列分析

Cloning and protein sequence analysis of alcohol dehydrogenase of Acetobacter pasteurianus AC2005

以具有优良醋酸发酵性能的巴斯德醋酸杆菌AC2005基因组DNA为模板,利用PCR的方法分别克隆了编码乙醇脱氢酶亚基Ⅰ和乙醇脱氢酶亚基Ⅱ的基因adhA和adhB。序列分析表明,adhA与GenBank已报道的序列(accessio nnumber:D13893.1)具有94%的同源性,氨基酸序列同源性达98%;adhB与GenBank已报道的序列(accession number:D13893.1)同源性为93%,氨基酸序列同源性达97%。利用TMHMM 2.0软件,对乙醇脱氢酶跨膜结构进行了分析,发现乙醇脱氢酶亚基Ⅰ和乙醇脱氢酶亚基II均为膜结合蛋白。利用Swiss-Model在线软件模拟了巴斯德醋酸杆菌AC2005乙醇脱氢酶亚基I的三维立体结构。该研究对该酶的结构和功能的进一步分析提供了基础。

The genes,adhA and adhB,coding the subunit I and II of alcohol dehydrogenase（ADH）were amplified by the polymerase chain reaction（PCR）,using genomic DNA of Acetobacter pasteurianus AC2005 as template,which had been demonstrated a potential strain for acetic acid production. The sequences were blasted in GenBank databases. The results showed that adhA shared 94% identity and 98% amino acid sequence homology,besides adhB shared 93% identity and 97% amino acid sequence homology with the reported gene（accession number：D13893. 1）. The characterization of transbilayer helix of ADH was analyzed,using software of TMHMM 2. 0. It was found that the subunits I and II were both membrane-bound protein. Furthermore,the three dimensional structure of subunit I of ADH in A. pasteurianus AC2005 was produced by using Swiss-Model workspace. Those results provided that some information for further researching the relationship of structure and function of ADH.