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《Journal of Huazhong Agricultural University(Natural Science Edition)》 2008-05
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Highly Effective Expression of Aspergillus niger Endo-β-1,4-glucanase in Pichia pastoris

HUANG Jun ZHANG Chang-yi ZHAO Shu-miao LIANG Yun-xiang(College of Life Science and Technology/ State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China)  
The gene encoding endo-β-1,4-glucanaseⅠ from Aspergillus niger L3 was isolated with RT-PCR.egⅠ was eliminated its native signal peptide by PCR and inserted into the pPIC9K vector of Pichia pastoris in the reading frame with α-factor secreting signal peptide sequence to construct the recombinant plasmid pPIC9k-egⅠ.The recombinant plasmid pPIC9k-egⅠ was transformed into P.pastoris GS115 with electroporation.Two of recombinant P.pastoris stains 1 and 5 were obtained by screen with MM,MD,CMC-Na and G418 plates and the activity of recombinant EGⅠ could reach to 1 456 U/mL and 1 928 U/mL,respectively.The optimal temperature for the recombinant EGⅠ was 70 ℃ and the optimal pH was 5.0.
【Fund】: 浙江省重大科技攻关项目(2004C12010)资助
【CateGory Index】: Q78
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