Reinigung von Kollagenase, III. Gewinnung enzymatisch einheitlicher Kollagenase

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Abstract
Crude, commercial collagenase, obtained by ammonium sulphate precipitation from culture filtrates of Clostridium histolyticum, contains pigments, enzymatically inactive proteins and peptides, several peptidases, esterases and amidase/-esterases. Carrier-free continuous electrophoresis, chromatogra-phy on polyamide powder and gel-filtration on Sephadex G-100 were combined to give a method for the removal of the contaminating enzymes. Collagenase, purified by the described procedure, showed an activity yield of 54% and a 39-fold purification over the starting material and was free from unspecific enzymic activities. The individual purification steps were followed by immunoelectrophoresis, using a rabbit antiserum prepared with crude collagenase. A method was devised for staining the collagenase In the specific immuno precipitates.