title: Purification of nonspecific protease-free collagenase from Clostridium histolyticum.
abstract: The collagenase (EC 3.4.24.3) produced by the bacterium Clostridium histolyticum has been purified free from nonspecific protease contaminants by a two-step procedure. The crude culture medium is chromatographed over heparin-Sepharose and Sephacryl S-200, and the resulting preparation has no activity versus noncollagenous proteins or N alpha-benzoyl-L-arginine ethyl ester, yet cleaves native thermally reconstituted collagen fibrils quite efficiently (specific activity, 3000 units/mg). The purification described may be useful for those investigators requiring substantially purified collagenase for applications such as cell culture or collagen quantitation in protein mixtures.
authors.forename: T A
authors.initials: TA
authors.lastname: Bicsak
authors.forename: E
authors.initials: E
authors.lastname: Harper
mesh_terms: D001120:Arginine / Q000031:analogs & derivatives / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D055598:Chemical Phenomena; D002621:Chemistry; D002845:Chromatography / Q000379:methods; D003013:Clostridium / Q000201:enzymology*; D003094:Collagen / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D003012:Microbial Collagenase / Q000302:isolation & purification*; D010447:Peptide Hydrolases / Q000302:isolation & purification; D013379:Substrate Specificity
chemical_list: D003094:Collagen; D001120:Arginine; C009727:benzoylarginine ethyl ester; D010447:Peptide Hydrolases; D003012:Microbial Collagenase