Semi-rational site-directed mutagenesis of phyI1s from Aspergillus niger 113 at two residue to improve its phytase activity

文献类型： 外文期刊

第一作者： Tian, Yong-Sheng

作者： Tian, Yong-Sheng;Peng, Ri-He;Xu, Jing;Zhao, Wei;Gao, Feng;Fu, Xiao-Yan;Xiong, Ai-Sheng;Yao, Quan-Hong

作者机构：

关键词： enzyme activity.;phyi1s gene;Phytase;Structure-function analysis;Internet resource.

期刊名称：MOLECULAR BIOLOGY REPORTS （ 影响因子：2.316； 五年影响因子：2.357 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Through alignment of amino acid sequences among different phytases, we found that the amino acid at residues 53 and 91 vary broadly. To prove that the amino acid at residues 53 and 91 were related to phytase specific activity, two single mutant phyI1s Q53R and K91D were obtained by site-directed mutagenesis strategy. None of the single amino acid residues in the two mutants was in a position reported to be important for catalysis or substrate binding. Kinetic analysis of the phytase activity of the two mutants (Q53R and K91D) indicated that the mutants were attributed to 2.2- and 1.5-fold increased specific activity, and a 1.47- and 1.16-fold increased affinity for sodium phytate. In addition, the overall catalytic efficiency (k cat/K m) of the two mutants was improved 4.08- and 2.84-fold compared to that of the wild type. Such mutants will be instrumental for the structure-function study of the enzyme and for industrial application.

分类号： Q7

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