Expression of a New Neutral Phytase in Escherichia coli and Its Purification and Enzymatic Characterization

Abstract

Abstract: A new neutral phytase gene (phyc) from Bacillus amyloliquefaciens DSM 1061 (GenBank: HM747163) was cloned into expression vector pET22b(+). The phytase was expressed in Escherichia coli BL21(DE3). The molecule weight of the phytase protein was about 42 kD determined by SDS-PAGE. The expressed phytase was about 30% of the total soluble protein of E. coli. The recombinant protein was purified by 6×His-Tagged Protein Purification Kit. Optimal pH value and temperature of the phytase were 7.0 and 60 ℃, respectively. Under the optimum conditions, the activity was 15 U/mg. The Km value of the phytase for hydrolysis of sodium phytate under 60 ℃ was 0.30 mmol/L.

Key words: Bacillus amyloliquefaciens, overexpression, neutral phytase, enzymatic properties

CLC Number:

Q814

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