Inhibition of Chymosin and the Coagulation of Para-Casein Micelles by Anions

Abstract

The effect of monovalent anions on the coagulation of para-casein micelles was determined by monitoring light transmission after adding different Ca salts (40 mM) to partially hydrolyzed casein micelles. The release of macro-peptide by chymosin was quantified using fluorescamine. The average rate of the chymosin-initiated coagulation of casein micelles, approximated by the reciprocal of clotting time, was determined as a function of anion type and concentration to determine the simultaneous effects of anions on chymosin velocity and para-casein micelle aggregability. Chymosin velocity and the coagulation of para-casein micelles were progressively inhibited by anions; the larger the anion, the greater the inhibition (SCN− > NO3− > Br− > Cl−). This is attributed to the binding of anions to cationic binding sites on kappa-casein and para-kappa-casein. The relative effect of the larger anions compared with that of Cl− on the average rate of the chymosin-initiated coagulation of casein micelles increased with anion concentration (6 to 120 mM) but became limited at 120 mM. In the presence of 120 mM SCN−, NO3− and Br−, the average rate of coagulation was 26, 59, and 78% of that obtained with 120 mM Cl−. The marked sensitivity of the interactions between chymosin and kappa-casein and between para-casein micelles to anion type support conclusions that cationic sites on kappa-casein are important in the mechanism of the chymosin-initiated coagulation of casein micelles. Thus, monovalent anions may be useful to elucidate the mechanism of protein-protein interactions in food systems. © 1986, American Dairy Science Association. All rights reserved.