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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; 37°C; 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 4°C in solutions with 0.05M CO32–-ions
and Co2+ ions
(10–5M).
Copyright (C) Chemistry Dept., Moscow State University, 2002
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