Purfication and Properties of Collegenase from a Streptomyces Species

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Abstract
A collagenase active against native, insoluble collagen was isolated from the culture fillrate of a streptomycete which has been designated Streptomyces sp. C-51. Collagenase was produced by growing this strain in media containing gelatin. Purification by ammonium sulfate fractionation and chromatographies on DEAE-Toyoperal and DEAE-Cellulofine columns produced active enzyme which was free of contaminating proteins, including nonspecific proteinases, but which contained two active subspecies (I and II). Both subspecies were purified by preparative slab gel electrophoresis. The apparent molecular weights were 100,000 for the homogeneous fraction I and 90,000–110,000 for the microheterogeneous fraction II. These two subspecies were most active at pH 8–9, very similar in amino acid composition and immunologically identical. Some other properties of the Streptomyces C-51 collagenase were copared with those of Clostridium histrolyticul collagenase. Substrate specificity, insensitivity to N-ethlmaleimide and diisopropyl fluorophosphate, and sensitivity to certain metal ion cmplexin agents were similar for the coollagenases from both microorganisms.