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High level expression of an acid-stable phytase from citrobacter freundii in pichia pastoris

文献类型： 外文期刊

作者： Zhao, Wei 1 ; Xiong, Aisheng 1 ; Fu, Xiaoyan 1 ; Gao, Feng 1 ; Tian, Yongsheng 1 ; Peng, Rihe 1 ;

作者机构： 1.Shanghai Acad Agr Sci, Biotechnol Res Inst, Shanghai 201106, Peoples R China

关键词： Citrobacter freundii;Codon optimization;pH-stable;Phytase;Pichia pastoris

期刊名称：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY （ 影响因子：2.926； 五年影响因子：2.685 ）

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收录情况： SCI

摘要： To obtain a high level expression of phytase with favorable characteristics, a codon-optimized phytase gene from Citrobacter freundii was synthesized and transferred into Pichia pastoris. Small-scale expression experiments and activity assays were used to screen positive colonies. After purified by Ni~(2+)-NTA agarose affinity column, the characterizations of the recombinant phytase were determined. The recombinant phytase (r-phyC) had two distinct pH optima at 2.5 and 4.5 and an optimal temperature at 50 °C. It retained more than 80% activity after being incubated under various buffer (pH 1.5-8.0) at 37 °C for 1 h. The specific activity, Km, and Vmax values of r-phyC for sodium phytate were 2,072±18 U mg~(-1), 0.52±0.04 mM, and 2,380±84 U mg~(-1) min~(-1), respectively. The enzyme activity was significantly improved by 1 mM of K ~+, Ca~(2+), and Mg~(2+). These characteristics contribute to its potential application in feed industry.

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