[Purification and study of some properties of a collagenase produced by Empedobacter collagenolyticum].
A collagenase from Empedobacter collagenolyticum was extracted from the culture medium of the bacteria. The complete purification of the enzyme was achieved by successive ammonium sulfate precipitation. Sephadex G 200 gel filtration and DEAE cellulose chromatography. This collagenase is active on insoluble collagen, and on the synthetic peptide Pz-Pro-Leu-Gly-Pro-D-Arg. Its optimum activity was at 30 degrees C and at pH 7.6. A strong inhibition was observed with chelating agents such as O-phenanthroline and EDTA. Among the cations tested to restore the activity, only Ca2+ has a measurable effect. Heavy metals, Pb, Hg, Cd, Cu, Fe, Co, strongly inhibit the enzyme activity. Zn2+ is also highly inhibitory; 10 microM  ZnCl2 completely inhibits the collagenase. p CMB, iodoacetate have little effect on the collagenase. This new collagenase ressembles by most of its properties the already known bacterial collagenases.
D001419:Bacteria / Q000201:enzymology*; D002118:Calcium / Q000494:pharmacology; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004492:Edetic Acid / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D003012:Microbial Collagenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D008970:Molecular Weight; D013379:Substrate Specificity
D004492:Edetic Acid; D003012:Microbial Collagenase; D002118:Calcium
Purification et étude de quelques propriétés d'une collagénase produite par Empedobacter collagenolyticum.