Renaturation of bacterial metalloproteinases
Vaganova, T.I.; Medvedeva, N.P.; Stepanov, V.M.
Biokhimiia 58(6): 913-920
1993
ISSN/ISBN: 0320-9725 PMID: 8364114 Document Number: 417498
After denaturation by phenol or acid ethanol bacterial metalloproteinases secreted by Bacillus thermoproteolyticus and Bacillus megaterium cells can be renatured by dissolution in 99.7% formic acid with subsequent dilution of the solution and its neutralization with an appropriate amount of an alkali. Renaturation is optimal at pH 9.0 in the presence of 30% glycerol as stabilizer, Ca2+ and Zn2+ ions needed for the formation of a native structure and reconstitution of the enzyme catalytic center. The active enzyme yield is 60-80%. Reactivated metalloproteinases retain their enzymatic properties, amino acid composition and molecular mass. Under these conditions autolysis of metalloproteinases does not significantly influence their renaturation.