title: Quantification of chymosin action on nonlabeled kappa-casein-related peptide substrates by ultraviolet spectrophotometry: description of kinetics by the analysis of progress curves.
abstract: A method is described for quantifying the proteolytic action of the milk-clotting enzyme chymosin on small and medium-sized peptide substrates by monitoring the decrease of absorbance at 230 nm during cleavage. The method is illustrated by the determination of the kinetic parameters of the specific splitting of a kappa-casein-related hexa- and pentadecapeptide by chymosin. The results are in good agreement with those found earlier with the same enzyme/substrate system by using an automated ninhydrin method. Erroneous results were obtained when the kinetic data were derived from one single progress curve. The significance of initial rate measurements for calculating correct kinetic parameters is briefly discussed. The usefulness of single progress curves measured at different initial substrate concentrations for obtaining information about the mechanism of the enzymic reaction is demonstrated.
mesh_terms: D000818:Animals; D002364:Caseins; D002417:Cattle; D012085:Chymosin / Q000032:analysis*; D007700:Kinetics; D009842:Oligopeptides; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity
chemical_list: D002364:Caseins; D009842:Oligopeptides; D012085:Chymosin