Kinetic characterization of affinity chromatography purified clostripain.

Abstract

The cysteine protease clostripain, purified by affinity chromatography on a large scale, shows very high activity against BAEE using the titrimetric standard assay. Furthermore, titration of the active site with the irreversible inhibitor tosyl-lysyl-chloromethane resulted in a more than two times higher specific activity compared with literature data (Porter et al. (1971) J. Biol. Chem. 246, 7675-7682). Based on the molar enzyme concentration determined, the hydrolysis kinetics of the standard substrate BAEE were compared with those for the N alpha-protected dipeptide ester Mal-Tyr-Arg-OEt. It was demonstrated from the kinetic data that the highly purified clostripain is the most active enzyme preparation available up to now. In contrast to the standard substrate, Mal-Tyr-Arg-OEt shows a threefold lower specificity constant.

Cite this paper

@article{Ullmann1994KineticCO, title={Kinetic characterization of affinity chromatography purified clostripain.}, author={Diana Ullmann and Hans Dieter Jakubke}, journal={Biological chemistry Hoppe-Seyler}, year={1994}, volume={375 2}, pages={89-92} }