Plasmin and coagulant activities in a minicurd model system: Study of technological parameters.

The effect of scalding temperature of the curd, the inclusion of a washing step, and the pH at whey drainage on plasmin and coagulant activities were assessed in a minicurd model of young hard cooked cheese. The variables were tested as follows: draining pH was assayed at 3 levels (4.6, 5.6, and 6.4), curd scalding temperature was tested at 50 and 56°C, and washing of the curd was examined at 2 levels (no washing step, and the replacement of the whey by water). Increase in pH at whey drainage and washing of the curd had a positive effect on plasmin activity, which was also evidenced by compatible changes in soluble peptide profiles. No effect of increased cooking temperature was found on plasmin activity. Plasminogen activation was not verified in any treatment. As for coagulant, lower pH values at whey drainage and a decrease in curd cooking temperature increased its activity; washing of the curd showed no influence on coagulant residual activity. These results were consistent with proteolysis described by peptide profiles, electrophoresis, and soluble nitrogen fractions.

Multivariate analysis of proteolysis patterns differentiated the impact of six strains of probiotic bacteria on a semi-hard cheese.

The individual contribution of 6 strains of probiotic bacteria (3 of Lactobacillus acidophilus and 3 of the Lactobacillus casei group) to proteolysis patterns in a semi-hard cheese was assessed. Control cheeses (without probiotics) and 2 types of experimental cheeses (with the addition of probiotics either directly to milk or by a 2-step fermentation method) were manufactured. Cheeses containing Lb. acidophilus showed the most extensive peptidolysis, which was evidenced by changes in the peptide profiles and a noticeable increase of free amino acids compared with control cheeses. The strains of the Lb. casei group showed a lower contribution to cheese peptidolysis, which consisted mainly of free amino acid increase. Two-step fermentation improved peptidolytic activity for only one of the cultures of Lb. acidophilus tested. The addition of Lb. acidophilus strains into cheese may be suitable not only for their beneficial health effect but also for their influence on secondary proteolysis, consistent with acceleration of ripening and improved flavor formation.

Influence of residual milk-clotting enzyme on alpha(s1) casein hydrolysis during ripening of Reggianito Argentino cheese.

Milk-clotting enzyme is considered largely denatured after the cooking step in hard cheeses. Nevertheless, typical hydrolysis products derived from rennet action on alpha(s1)-casein have been detected during the ripening of hard cheeses. The aim of the present work was to investigate the influence of residual milk-clotting enzyme on alpha(s1)-casein hydrolysis in Reggianito cheeses. For that purpose, we studied the influence of cooking temperature (45, 52, and 60 degrees C) on milk-clotting enzyme residual activity and alpha(s1)-casein hydrolysis during ripening. Milk-clotting enzyme residual activity in cheeses was assessed using a chromatographic method, and the hydrolysis of alpha(s1)-casein was determined by electrophoresis and high performance liquid chromatography. Milk-clotting enzyme activity was very low or undetectable in 60 degrees C- and 52 degrees C-cooked cheeses at the beginning of the ripening, but it increased afterwards, particularly in 52 degrees C-cooked cheeses. Cheese curds that were cooked at 45 degrees C had higher initial milk clotting activity, but also in this case, there was a later increase. Hydrolysis of alpha(s1)-casein was detected early in cheeses made at 45 degrees C, and later in those made at higher temperatures. The peptide alpha(s1)-I was not detected in 60 degrees C-cooked cheeses. The results suggest that residual milk-clotting enzyme can contribute to proteolysis during ripening of hard cheeses, because it probably renatures partially after the cooking step. Consequently, the production of peptides derived from alpha(s1)-casein in hard cheeses may be at least, partially due to this proteolytic agent.

Influence of milk-clotting enzyme concentration on the alphas1-casein hydrolysis during soft cheeses ripening.

We studied the influence of the dose of milk-clotting enzyme on alphas1-CN degradation, soluble nitrogen production, and sensory profile for an Argentinean soft cheese: Cremoso Argentino. Five different types of cheeses were produced: 1) control cheeses with normal technology, 2) cheeses with inactivated milk-clotting enzyme, 3) cheeses with inactivated milk-clotting enzyme, without starter (acidified with glucono delta lactone), 4) cheeses with a half dose of milk-clotting enzyme, and 5) cheeses with a double dose of milk-clotting enzyme. Proteolysis was assessed by isoelectric focusing electrophoresis of the insoluble fraction at pH 4.6, followed by densitometric quantification. Soluble nitrogen at pH 4.6, expressed as a percentage of total nitrogen and defined as ripening index was also performed. A sensorial panel evaluated the cheeses at the end of ripening. The hydrolysis level of alphas1-CN depended on the milk-clotting enzyme dose used in cheese making. Cheeses without active coagulant did not show degradation at the end of ripening, while cheeses with half and whole doses showed proportional degradations to coagulant dose. Cheese with a double dose of coagulant did not show higher alphas1-CN hydrolysis than normal cheese. No difference was found between cheeses with and without microbiological starter, indicating that the selected culture, composed of thermophilic strains, was unable to attack the whole casein. A high linear correlation was found between ripening index and the relation Sensorial characteristics of cheeses agree with objective analysis. Cheeses without active coagulant were hard and crumbly, while cheeses with normal dose were soft and creamy.