Gene Cloning, Expression, and Characterization of a Novel Phytase from Dickeya paradisiaca

文献类型： 外文期刊

第一作者： Gu, Weina

作者： Gu, Weina;Huang, Huoqing;Meng, Kun;Yang, Peilong;Fu, Dawei;Luo, Huiying;Wang, Yaru;Yao, Bin;Gu, Weina;Zhan, Zhichun

作者机构：

关键词： Degenerate PCR;Dickeya paradisiaca Histidine acid phosphatase;Phytase;TAIL-PCR

期刊名称：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY （ 影响因子：2.926； 五年影响因子：2.685 ）

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年卷期：

页码：

收录情况： SCI

摘要： A novel phytase gene, appA, was isolated by degenerate polymerase chain reaction (PCR) and thermal asymmetric interlaced PCR from Dickeya paradisiaca. The full-length appA comprises 1278 bp and encodes 425 amino acid residues, including a 23-residue putative N-terminal signal peptide. The deduced amino acid sequence of appA reveals the conserved motifs RHGXRXP and HD, which are typical of histidine acid phosphatases; significantly, APPA shows maximum identity (49%) to a phytase from Klebsiella pneumoniae. To characterize the properties of APPA, appA was expressed in Escherichia coli and purified. The purified recombinant APPA has two pH optima at pH 4.5 and 5.5, optimum temperature at 55 °C, specific activity of 769 U/mg, and good pH stability. The Km value for the substrate sodium phytate is 0.399 mM with a Vmax of 666 U/mg. To our knowledge, this is the first report of a phytase or phytase gene isolated from Dickeya.

分类号： Q5

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