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利用随机突变探讨影响α—淀粉酶催化活性的氨基酸位点 被引量:5
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作者 缪可嘉 张大龙 +1 位作者 马延和 王正祥 《食品工业科技》 CAS CSCD 北大核心 2007年第10期63-65,69,共4页
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 i... 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. 展开更多
关键词 Α-淀粉酶 活性位点 随机突变 空间构象 氢键
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酿酒酵母表面展示β-淀粉酶及其酶学性质研究 被引量:2
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作者 杨华 樊游 +3 位作者 沈微 石贵阳 SurenSingh 王正祥 《食品工业科技》 CAS CSCD 北大核心 2012年第13期135-138,共4页
通过PCR技术扩增来源于大麦的β-淀粉酶基因,将其与酿酒酵母细胞壁蛋白α凝集素基因在读框内融合,构建得到表面展示载体pBA-AG,进一步将该重组质粒通过遗传转化,整合到酿酒酵母W303-1A的染色体中,获得了β-淀粉酶经过α凝集素锚定信号... 通过PCR技术扩增来源于大麦的β-淀粉酶基因,将其与酿酒酵母细胞壁蛋白α凝集素基因在读框内融合,构建得到表面展示载体pBA-AG,进一步将该重组质粒通过遗传转化,整合到酿酒酵母W303-1A的染色体中,获得了β-淀粉酶经过α凝集素锚定信号结合到细胞壁上的重组酵母。重组酵母表面展示的β-淀粉酶活力为131U/g干细胞。对展示的β-淀粉酶酶学性质研究表明,其最适反应温度为50℃,最适作用pH为5.0,与游离酶相比,其温度稳定性和pH稳定性均得到提高。本研究利用α凝集素系统首次将β-淀粉酶成功展示在酿酒酵母表面,为以酿酒酵母为基础的全细胞催化剂研究与应用打下了一定基础。 展开更多
关键词 酿酒酵母 Β-淀粉酶 表面展示 全细胞催化
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