RENATURATION OF BACTERIAL METALLOPROTEINASES

Bacterial metalloproteinases secreted by Bacillus thermoproteolyticus and Bacillus megaterium which have been denatured by phenol or acidic ethanol can be renatured by dissolving them in 99.7% formic acid, dilu tion, and neutralization of the solution with the calculated amount of alkali. The renaturation is most effective at pH 9.0 in the presence of 30% glycerol as a stabilizer and calcium and zinc ions which are ne cessary for the formation of the native structure and rebuilding of th e catalytic center of the enzymes. The yield of active enzymes is 60-8 0%. The reactivated metalloproteinases retain their enzymatic properti es, amino acid composition, and molecular weight. The autolysis of the metalloproteinases does not affect the renaturation under these condi tions.