SUMMARY. Casein phosphopeptides (CPP) were produced by tryptic hydrolysis of sodium caseinate and further purified by precipitation and chromatography on QAE-Sephadex A-25. Their physico-chemical properties were compared with the properties of an enzymically dephosphorylated equivalent preparation (DPP). Binding of Ca 2+ to the peptides was measured using a Ca selective electrode and was found to increase with pH and to show 1/1 stoicheiometry Ca/P org in CPP at pH 6 -5 a.nd 7-6. Klotz plots indicated equivalent binding sites at these two pH values, but some heterogeneity was seen at pH 3-5. In contrast, DPP did not bind significant amounts of Ca 2+ . CPP effectively inhibited the formation of insoluble calcium phosphates at different Ca/P ratios. The effective CPP concentration was 10 mg/1 and complete stability of calcium phosphate solutions was obtained at about 100 mg/1. This stabilizing effect was dependent on the presence of organic P.

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The conditions under which a high volume fraction of oil can be trapped in whey protein gels were studied. Oil-in-water emulsions of whey protein and vegetable oil were subjected to heat treatment. Such emulsions, depending on their protein and oil content, on their pH and on the emulsification technique used, gelled or remained liquid. Homogenization was the major factor to achieve gelation and the firmness of heat-induced gels increased with increasing emulsion fineness and homogeneity. Emulsions with a high gelation capacity were characterized by a single droplet family of relatively narrow size distribution and a mean droplet diameter ranging from roughly 300-700 nanometers. The pH range suitable for gelation extended from 3.5-8.0.

The specific IgE binding capacity of native bovine α-lactalbumin (α-La), a globular whey protein, and tryptic peptides was investigated using 19 sera from patients with cow’s milk protein allergy. The specific anti-bovine α-La IgE titers ranged from 0.6 to 125 IU/ml. Highly purified tryptic peptides from native and disulfide-bond-reduced α-La were obtained by reverse phase chromatography. By ELISA technique using immobilized native protein or peptides, 11 of the 19 sera reacted exclusively with intact protein while 8 of them also presented a specific IgE response to different tryptic peptides. Polyclonal IgE population specificity was not related to anti-bovine α-La IgE levels. Sequence (17G-K58) and larger peptides sharing this sequence were most strongly and frequently recognized. Competitive ELISA inhibition tests confirmed this IgE-specific response and gave also clear evidence for IgE binding to smaller peptides corresponding to sequences (6C-R10):S-S:(115L-L123) and (109A-L123). IgE binding to native α-La and large peptides confirmed the importance of conformational epitope(s). However, in some sera reduced and S-alkylated peptide (59I-K94) exhibited a similar or higher IgE binding capacity than the native corresponding fragment, suggesting the existence of sequential epitope(s) exposed through protein denaturation. Moreover, IgE binding sequences were also located within hydrophobic regions of α-La and/or within parts with high sequence homology to human α-La.

Partial enzymatic hydrolysis of whey protein by trypsin increased solubility of this protein in water. Water-insoluble, heat-denaturated whey protein was solubilized fully by trypsinization. Optimal conditions for the enzyme reaction, established by the pH-stat technique, were: digestion at pH 8.0 and 55 C for approximately 3 h, at an enzyme-substrate ratio of 1 : 100. Under these conditions, 500 mumoles of titratable protons were liberated per g of substrate in the course of the reaction. Digestion at 40 C generated only about 400 mumoles of acid. Predenaturation of the substrate by heat did not improve digestibility. The extent of hydrolysis reached approximately 8% of all peptide bonds in the protein. Fractionation of the digest on Sephadex G-50 showed it was composed of a major fraction of highly water soluble peptides, ranging in molecular weight from approximately 500 to 5000. The gel excluded a minor fraction of larger, aggregated peptides. This aggregate was dissociated in the presence of urea and a reducing agent. All amino acids in the digest, except some lysine and arginine, were peptide bound.