[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.