Studies on collagenase in fish. II. Some properties of a collagenase from the pyloric caeca of Seriola quinqueradiata.

Abstract

A collagenolytic enzyme has been isolated from the pyloric caeca of yellow-tail, Seriola quinqueradiata, The pyloric caecum collagenase acts on native collagen fibrils and on collagen in solution. The activity is maximal at pH 8. 5. Diisopropyl fluoro-phosphate, N-tosyl-L-lysylchloromethane, soybean trypsin inhibitor, and human and yellow-tail sera inhibit this enzyme; EDTA and N-tosyl-L-phenylalanylchloromethane are less inhibitory. This enzyme is somewhat activated by cysteine while such sulfhydryl reagents as p-chloromercuribezoate, monoiodoacetate, and N-ethylmaleimide have no effect. This suggests that pyloric caecum collagenase does not require sulfhydryl groups for activity even though it is slightly activated by cysteine. This enzyme activity, as investigated by polyacrylamide disc electrophoresis, results in a marked decrease in the original β-component and the apearance of numerous new components beneath the original α-band. Its mode of attack on collagen and sensitivity to various inhibitors indicate that this enzyme resembles crab hepatopancreas collagenase rather than most known animal collagenases.