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ABSTRACT: Hydrolysis-induced coagulation of casein micelles by pepsin occurs during the digestion of milk. In this study, the effect of pH (6.7–5.3) and pepsin concentration (0.110–2.75 U/mL) on the hydrolysis of κ-casein and the coagulation of the casein micelles in bovine skim milk was investigated at 37°C using reverse-phase HPLC, oscillatory rheology, and confocal laser scanning microscopy. The hydrolysis of κ-casein followed a combined kinetic model of first-order hydrolysis and putative pepsin denaturation. The hydrolysis rate increased with increasing pepsin concentration at a given pH, was pH dependent, and reached a maximum at pH ∼6.0. Both the increase in pepsin concentration and decrease in pH resulted in a shorter coagulation time. The extent of κ-casein hydrolysis required for coagulation was independent of the pepsin concentration at a given pH and, because of the lower electrostatic repulsion between para-casein micelles at lower pH, decreased markedly from ∼73% to ∼33% when pH decreased from 6.3 to 5.3. In addition, the rheological properties and the microstructures of the coagulum were markedly affected by the pH and the pepsin concentration. The knowledge obtained from this study provides further understanding on the mechanism of milk coagulation, occurring at the initial stage of transiting into gastric conditions with high pH and low pepsin concentration.

(+)-Catechin, the representative catechin in green tea, was incorporated into a full-fat cheese (at 125–500 ppm) followed by ripening for 90 days at 8 °C and digesting for six hours. Determination of pH, proximate composition, total phenolic content (TPC) and antioxidant activity (AA) after manufacture and ripening demonstrated that the addition of (+)-catechin significantly (p ≤ 0.05) decreased the pH of both whey and curd during cheese manufacturing and ripening with no significant (p > 0.05) effect on the moisture, protein and fat contents. (+)-Catechin increased TPC, as well as AA, though the increase was not proportional with increasing the concentration of added (+)-catechin. About 57%–69% of (+)-catechin was retained in the cheese curd, whereas about 19%–39% (depending on the concentration) was recovered from the cheese digesta. Transmission electron micrographs showed that the ripened control cheese had a homogeneous pattern of milk fat globules with regular spacing entrapped in a homogenous structure of casein proteins, whereas the addition of (+)-catechin disrupted this homogenous structure. The apparent interaction between (+)-catechin and cheese fat globules was confirmed by Fourier transform infrared spectroscopy. These associations should be taken into account when incorporating antioxidants, such as (+)-catechin, to create functional dairy products, such as cheese.