Camembert cheeses were made with pasteurized skim milk or with ultrafiltered milk concentrated 2-fold, using either rennet or Mucor miehei proteinase as coagulant. Using rennet, the rapid acidification during cheesemaking increased enzyme retention and 55% was retained after 24 h. With the M. miehei proteinase, acidification had no effect and only 17% of it was retained after 24 h. The use of ultrafiltered milk resulted in less rennet being retained in the curd.

A method for determining chymosin and bovine pepsin A in commercial extracts of bovine veils, based on the use of the synthetic hexapeptide (Leu-Ser-Phe(NO2)-Nle-Ala-Leu-OMe) as reference substrate, is reported. Chymosin and bovine pepsin A were separated chromatographically from extracts and assayed for clotting activity on a reconstituted skim-milk standardized with reference chymosin and bovine pepsin A, themselves standardized in relation to the hexapeptide. The effect of pH on the absorbance difference between the hexapeptide and the Leu-Ser-Phe(NO2) tripeptide resulting from its hydrolysis was studied. It was found that the ‘optimal’ pH for determining the activities of the reference enzyme solutions was 4·7.

Six chymosin and 3 bovine pepsin A preparations were assayed on the hexapeptide to define the relation between the proteolytic activity and the amount of active enzyme. At pH 4·7 and 30 °C 1 mg chymosin and 1 mg bovine pepsin A hydrolysed 100 and 2700 μm-peptide/s respectively. The clotting activity of these preparations was assayed on a reconstituted skim-milk to standardize it and thus define the relation between the clotting time and the amount of active enzyme. Chymosin had a specific clotting activity twice that of bovine pepsin A. At equal clotting activities, bovine pepsin A was 55 times more active than chymosin on the hexapeptide at pH 4·7.

Two stages of gelling by the milk-clotting enzymes chymosin and pepsin were studied at different milk protein concentrations, using milk retentates. In each case, enzymic velocity versus protein concentration described a standard hyperbola. According to the experimental conditions such as pH and type of enzyme, which changed the enzyme concentration used, either the quasi-linear part of the hyperbola was observed, or the velocity hardly increased and tended to a limiting value. Gelling occurred with a lower degree of proteolysis of κ-casein when the protein concentration increased but a minimum proteolysis (1% of total nitrogenous matter content) was required for aggregation to take place. Gelling time varied with the protein concentration, the pH and the enzyme concentration. The final degree of proteolysis of κ-casein was the same whatever the substrate concentration used.

Studies were undertaken to determine the influence of dietary proteins on the rennin and pepsin contents of preruminant calf vell. Three groups of 12 Friesian calves were each fed either milk proteins, whey proteins or a 50:50 mixture of these 2 diets. They were slaughtered at a constant weight of 150kg and their vells collected and dried. Another group of vells was obtained from 8 animals that had been fed milk proteins in a separate experiment. The extraction of the abomasal enzymes was carried out at acid pH, and the extracts were quantitatively analysed for rennin and pepsin by DEAE-cellulose column chromatography. Qualitative analyses were also performed by agarose-acrylamide gel electrophoresis. The only enzymes observed using this last method were rennin and bovine pepsin II. Statistical analysis of the quantitative enzyme determinations indicated a trend for the vells from calves fed diets containing casein to be richer in total activity and in rennin, while the level of pepsin remained approximately constant. It seems that casein may induce the secretion of rennin. However, further experiments will be necessary to confirm this. Important differences were observed between the 2 groups of veils from calves given the same diet, but grown in slightly different conditions.