[Purification and physico-chemical properties of collagenase synthesized by a bacterium of the type Acinetobacter sp].
The Acinetobacter spec collagenase has been almost completely purified. This enzyme is a true collagenase the activity of which is high on collagen. The enzyme is active on insoluble collagen, gelatin and the synthetic Pz-peptide, but has no proteolytic activity on casein or bovine serum-albumin. The collagenase was obtained on a simple medium with gelatin and yeast extract. The enzyme was purified by (NH4)2SO4 precipitation. DEAE cellulose column chromatography, Sephadex G 200 gel-filtration. The molecular weight of the enzyme was found to be 102 000 daltons, and its isoelectric point was found to be 7,7 +/- 0,2. The optimum pH and temperature for insoluble collagen hydrolysis were 7.6 and 37 degrees C, respectively; so, this collagenase corresponds to true collagenase. Hydrolysis of Pz-peptide is activated by Ca2+ and inhibited by metal ions (Cu2+, Fe3+, Zn2+, Pb2+, Hg2+). EDTA and o-phenanthroline induced a very significant reduction in enzyme activity. Iodoacetate and p-CMB induced a slight reduction in enzyme activity only at high concentrations (10-2M). The collagenase is most stable for temperatures less than or equal to 50 degrees C.
D000150:Acinetobacter / Q000201:enzymology*; D002413:Cations, Divalent; D004492:Edetic Acid / Q000494:pharmacology; D004789:Enzyme Activation; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000494:pharmacology; D007700:Kinetics; D003012:Microbial Collagenase / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D010618:Phenanthrolines / Q000494:pharmacology
D002413:Cations, Divalent; D010618:Phenanthrolines; D004492:Edetic Acid; D007501:Iron; D003012:Microbial Collagenase
Purification et propriétés physico-chimiques d'une collagénase synthétisée par une bactérie du genre Acinetobacter sp.