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D. A. Gudkov, E. N. Efremenko

The Refolding of Organophosphorous Hydrolase Containing the Hexahistidine Tag from Inclusion Bodies

Abstract

The inclusion bodies of organophosphorous hydrolase with hexahistidine tag at the N-terminus of protein molecule were isolated from E.coli DH5 cells and purified. Optimal conditions for solubilization of the inclusion bodies, as it was established, were following: 6 M urea in phosphate-saline buffer with pH 7.6; 37C; 2h. The refolding of the enzyme from solutions of solubilized inclusion bodies was carried out using metal-chelating chromatography. The enzyme activation after its refolding was studied. It was shown that maximum of catalytic activity of the enzyme can be obtained after 24h-incubation at 4C in solutions with 0.05M CO32-ions and Co2+ ions (105M).
Moscow University Chemistry Bulletin.
2007, Vol. 48, No. 6, P. 389
   

Copyright (C) Chemistry Dept., Moscow State University, 2002
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