Expression and Characterization of Aquatic Neutral Phytase Gene from Penicillium sp. C1 in Pichia pastoris

doi:10.13560/j.cnki.biotech.bull.1985.2019-0704

Abstract

Abstract: A new phytase gene was cloned from the genome DNA of Penicillium sp. C1 strain by degenerate primers and TAIL-PCR,and named as phyC1. The whole length of structural gene was 1 477 bp,including a 58 bp intron at the 5' terminus. The gene encoded 472 amino acids and a termination codon,and the first 23 amino acids were signal peptides. The gene encoding the mature protein of phyC1 was efficiently expressed in Pichia pastoris GS115,and then the recombinant enzyme PhyC1 was purified to electrophoretic purity by ammonium sulfate precipitation,anion exchange column and Sephedex chromatography. The enzymatic properties showed that the optimal pH of recombinant PhyC1 was 6.5,and high stability at pH 5.0-10.0. The optimal temperature of recombinant PhyC1 was 50℃. Compared with the commercial phytases,PhyC1 showed two significant properties,such as high activity under low temperature,especially below 20℃,and over 90% relative enzymatic activity remained under neutral pH 7.0. Therefore,PhyC1 has potential application prospect in feed industry,especially in aquatic industry.

Key words: neutral phytase, aquatic enzymes, Penicillium, Pichia pastoris

LI Ya-nan, YU Li-hong, CHEN Xin-mei, YANG Hao-meng, HUANG Huo-qing. Expression and Characterization of Aquatic Neutral Phytase Gene from Penicillium sp. C1 in Pichia pastoris[J]. Biotechnology Bulletin, 2020, 36(2): 134-141.

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