Overexpression and characterization of appA phytase expressed by recombinant baculovirus-infected silkworm

文献类型： 外文期刊

第一作者： Yin, C

作者： Yin, C;Zhu, ZZ;Lin, XA;Yi, YZ;Zhang, ZF;Shen, GF

作者机构：

关键词： Escherichia coli;appA;phytase;baculovirus;silkworm

期刊名称：JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：2.351； 五年影响因子：2.65 ）

ISSN： 1017-7825

年卷期： 2005 年 15 卷 3 期

页码：

收录情况： SCI

摘要： An Escherichia coli strain with high phytase activity was screened from pig excreta. The phytase gene, appA, was amplified by PCR technique. To obtain large amounts of appA phytase, the appA gene was subcloned into the baculovirus transfer vector pVL 1393 under the control of the Polyhedrin promoter. The recombinant baculovirus harboring the appA gene was obtained after co-transfection and screening. The early 5(th) instar larvae of silkworm were infected with the recombinant virus. Using this system, the appA phytase was overproduced up to 7,710 U per ml hemolymph. SDS-PAGE analysis revealed the baculovirus-derived appA phytase to be approximately 47 kDa in size. The optimal temperature and pH of the expressed phytase were 60 degrees C and pH 4.5, respectively. The enzymatic activity was increased by the presence of 1 mM Ca2+, 1 mM Mn2+, or 0.02% Triton X-100.

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