DOI: 10.3724/SP.J.1145.2009.00371

Chinese Journal of Appplied Environmental Biology (应用与环境生物学报) 2009/15:3 PP.371-375

Characteristics of Phytase from Escherichia coli*

Gene appA was cloned into prokaryotic expression vector pET32a to construct pET32a-appA and gene appA was successfully expressed in Escherichia coli Origami (DE3). Recombinant phytase was purified and its enzymatic characterization were studied. The results of SDS-PAGE showed higher purity of the enzyme after purified by nickel-chelating sepharose chromatography. Its specific activity was 3.36×106 U/mg, and optimal pH and temperature for its activity were 4.5 and 60 ℃, respectively. The dry thermal stability of phytase was stronger than wet thermal stability at 80 ℃ and 85 ℃. The enzyme activity was activated by Mg2+, Ca2+ and Mn2+ , and inhibited by Co2+, Cu2+ and K+. However, it almost lost activity when influenced by Fe3+ and Zn2+. In addition, it had strong ability to resist hydrolyzation by pepsin and partial ability to resist hydrolyzation by trypsin. Fig 8, Tab 1, Ref 21

Key words:appA-encoded phytase,prokaryotic expression;nickel-chelating sepharose chromatography,enzymatic characterization

ReleaseDate:2014-07-21 15:02:21

Funds:Supported by the Key Project of the Department of Science and Technology of Sichuan, China (No. 07GG111-03)

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