Calcium-binding Peptides Derived from Tryptic Hydrolysates of Cheese Whey Protein

Substrates and enzymesFresh Holstein milk was obtained from a Gyeongsang National University livestock farm.Cheddar cheese was manufactured according to the methods of Kosikowski (1982), and CWP separated from the cheese process contained 1.03% protein, 4.26% lactose, 0.7% ash and 93.53% moisture (determined by AOAC method, 1990).Defatting and demineralization of CWP were performed by ultracentrifuge (Supra 25 K, Hanil Sci., Korea) and dialysis (Sigma Co., USA), respectively.CWP was adjusted to pH 8.0 through the addition of 0.5 N NaOH and then heattreated for 10 min at 100°C.Trypsin (Bovine pancreas, activity 3.3 Anson units/g protein) was purchased from Novo Nordisk A/S (Bagsvaerd, Denmark). Tryptic hydrolysis and selective precipitation of calcium-binding peptidesHydrolysis of heated-CWP was determined by the Calcium-binding Peptides Derived from Tryptic Hydrolysates of Cheese Whey Protein