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High expression of a heat-stable phytase in Pichia pastoris

文献类型： 外文期刊

作者： Peng, RH 1 ; Xiong, AS 2 ; Li, X 2 ; Fan, HQ 2 ; Yao, QH 2 ; Guo, MJ 2 ; Zhang, SL 2 ;

作者机构： 1.Shanghai Acad Agr Sci, Agro Biotechnol Res Ctr, Shanghai Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China

2.Shanghai Acad Agr Sci, Agro Biotechnol Res Ctr, Shanghai Key Lab Agr Genet & Breeding, Shanghai 201106, Peoples R China; E China Univ Sci & Technol, Dept Bioengn, Shanghai 200237, Peoples R China

关键词： heat-stable phytase;gene synthesis;high expression;Pichia pastoris

期刊名称：ACTA BIOCHIMICA ET BIOPHYSICA SINICA （ 影响因子：3.848； 五年影响因子：3.67 ）

ISSN： 0582-9879

年卷期： 2002 年 34 卷 6 期

页码：

收录情况： SCI

摘要： It is difficult to obtain naturally occurring phytase having the required thermostability for application in animal feeding. The 1.3 kb thermal stable phytase gene (fphy) of Aspergillus fumigatus was synthesized by using a successive PCR method for the optimal expression in Pichia pastoris. Though the nucleotides of synthetic fphy share only 74% homology with the natural gene, the amino acid sequences coded by both genes were identical. After being cloned into the yeast expression vector (pPIC9) and inserted into the chromosome of Pichia pastoris by homologous recombination, phytase was expressed in the yeast and secreted from the cell. The strains for phytase over-expression were selected out by SDS-PAGE and enzyme analysis. After fermentation in 5 L fermention tank and induced by 0.5% methanol for 60 h, about 5.6 g purified phytase was obtained per liter culture fluid. The activity of phytase was 130 000 a per microlitre fluid. The thermostable phytase remained 40% active after exposure at 90 degreesC for 80 min.

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[1]Influence of signal peptide sequences on the expression of heterogeneous proteins in Pichia pastoris. Xiong, AS,Peng, RH,Li, X,Fan, HQ,Yao, QH,Guo, MJ,Zhang, SL. 2003

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