Gene Cloning，Expression and Characterization of an HAP Phytase from Pseudomonas fluorescens 206

Abstract

Abstract: A Pseudomonas fluorescens 206 strain producing HAP phytase was isolated from the bottom mud of artificial fish ponds in Jiaxin city. A novel HAP phytase encoding gene phyP was cloned by degenerate PCR and TAIL-PCR. The full length gene consist 1 284 bp and encodes 427 amino acids, including a putative signal peptide of 24 residues. The gene phyP was expressed in E.coli. The recombinant enzyme PhyP was purified to homogeneity by ammonium sulfalte precipitation and anion exchange chromatography. The optimal pH of recombinant PhyP was pH5.5, and the enzyme showed high stability at pH3.5-7.5. The optimal temperature of PhyP was found to be 45 ℃, and the enzyme had a higher activity at 25 ℃. These superior properties of PhyP make it advantageous for applying in feed industries, especially fisher industry.

Key words: Pseudomonas fluorescens, HAP phytase, Gene cloning and expression, Characteristics

Li Yanan, Huang Huoqing, Yao Bin, Zhao Qing, Liu Kun, Ma Wenkang. Gene Cloning，Expression and Characterization of an HAP Phytase from Pseudomonas fluorescens 206[J]. Biotechnology Bulletin, 2013, 0(8): 139-144.

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