利用随机突变探讨影响α—淀粉酶催化活性的氨基酸位点  

作　　者：缪可嘉[1] 张大龙[1] 马延和[1] 王正祥[1] 

机构地区：[1]江南大学工业生物技术教育部重点实验室和江南大学生物工程学院,江苏无锡214036

出　　处：《食品工业科技》2007年第10期63-65,69,共4页Science and Technology of Food Industry

基　　金：新世纪优秀人才支持计划(NCET-04-9704);国家高技术发展计划(2006AA020204)资助

摘　　要：α-Amylase is widely used in biotechnology industries such as food,textile and detergent.It occupies an important position in the domestic and international enzyme market.An important reason that amylase can achieve its function is it has catalysis domain,which is a very basilic domain,and the function of the catalysis domain mainly depends on catalytic active site.So changing the base of the active site or the base around the active site may affect a lot on α-amylase.This article mainly describes using random mutagenesis of the α-amylase by error-prone PCR,detecting the amylase activity of the mutant strains,and then sequencing the mutants.8 available mutants was picked to do further analysis and prediction of 3D structure.Choosing the mutant with the highest enzyme activity improvement as a further research target molecule,the potential structure changes were predicted by replacement of amino acid residue at the mutant site with other 18 residues.α-Amylase is widely used in biotechnology industries such as food, textile and detergent. It occupies an important position in the domestic and international enzyme market. An important reason that amylase can achieve its function is it has catalysis domain, which is a very basilic domain, and the function of the catalysis domain mainly depends on catalytic active site. So changing the base of the active site or the base around the active site may affect a lot on α- amylase. This article mainly describes using random mutagenesis of the α-amylase by error-prone PCR, detecting the amylase activity of the mutant strains, and then sequencing the mutants. 8 available mutants was picked to do further analysis and prediction of 3D structure. Choosing the mutant with the highest enzyme activity improvement as a further research target molecule, the potential structure changes were predicted by replacement of amino acid residue at the mutant site with other 18 residues.

关 键 词：Α-淀粉酶 活性位点 随机突变 空间构象 氢键 

分 类 号：TS202.25[轻工技术与工程—食品科学]