Bovine beta-casein was purified from phosphocasein by rennet coagulation and cold solubilisation from the resultant curd. beta-Casein was then dephosphorylated using potato acid phosphatase. Urea-polyacrylamide gel electrophoresis (PAGE) of partially dephosphorylated beta-casein showed a number of bands, depending on the final level of phosphorylation. Dephosphorylating beta-casein increased its pH of minimum solubility from similar to pH 5 to 5.5 and reduced its net negative charge from -30.8 to -27.0 mV. During the acidification of beta-casein solutions, partially dephosphorylated beta-casein failed to form a gel, unlike the phosphorylated (i.e., control) beta-casein. Use of partially dephosphorylated (beta-casein to stabilise oil-in-water emulsions resulted in larger fat globules compared to control beta-casein, but such globules were less susceptible to aggregation in the presence of 15 or 30 mM CaCl2. Overall, the dephosphorylation of beta-casein resulted in a protein similar to human beta-casein in terms of physicochemical functionality, with increased stability against calcium-induced aggregation. (C) 2012 Elsevier Ltd. All rights reserved.