Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Expression of the Aspergillus niger var. awamori Phytase Gene in Pichia pastoris, and Comparison of Biological Properties

  • CHOI, JAE-MUN (Graduate School of Biotechnology, Korea University) ;
  • DOO-SANG KIM (Department of Food Science & Biotechnology, Pukyung National University) ;
  • MOON-SICK YANG (Institute for Molecular Biology and Genetics, Chonbuk National Univesity) ;
  • HYUNG-RAK KIM (Department of Food Science & Biotechnology, Pukyung National University) ;
  • JAE-HO KIM (Graduate School of Biotechnology, Korea University)
  • Published : 2001.12.01
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Abstract

The PhyA gene, encoding myo-inositol hexakisphosphate phosphohydrolase in Aspergillus niger var. awamori (wild-type), was cloned and sequenced. The cDNA was overexpressed by a multicopy gene expression system in Pichia pastoris KM71. Recombinant, wild-type and commercial phytase from Aspergilus ficuum NRRL 3135 (Natuphos) were purified. The PhyA gene of Aspergillus niger var awamori showed perfect homology to the phytase of Aspergillus ficcum and $97\%$ homology to A. niger var awamori (L02421). Wild-type phytase was highly glycosylated and more thermostable than the other two, while deglycosylated farms of three phytases showed identical molecular weight, 507 kDa. After heating at $80^{\circ}C$, wild-type, commercial, and recombinant phytases retained $57\%, 32%,\;and\;8\%$ of their original activities, respectively. In conclusion, glycosylation plays a key role in the thermostability of phytase and its enzymatic characterization.

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