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Evaluation of the Residual Antigenicity of Dairy Whey Hydrolysates Obtained by Combination of Enzymatic Hydrolysis and High-Pressure Treatment

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Abstract:

Dairy whey was hydrolyzed for 15 min with five food-grade enzymes (Alcalase, Neutrase, Corolase 7089, Corolase PN-L, and Papain) at atmospheric pressure (0.1 MPa) and in combination with high pressure (HP) at 100, 200, and 300 MPa, applied prior to or during enzymatic digestion. The peptide profile of the hydrolysates obtained was analyzed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), and their residual antigenicity was assessed by immunoblotting with anti–β-lactoglobulin monoclonal antibodies and the sera from pediatric patients allergic to cow's milk proteins. Moreover, to evaluate the presence of residual trace amounts of casein in bovine whey hydrolysates, immunoblotting with anti–cow's milk protein polyclonal antibodies was performed. SDS-PAGE analysis showed that HP treatment increased hydrolysis by the proteases assayed, especially when it was applied during the enzymatic digestion. Positive reactions at the band corresponding to β-lactoglobulin were detected for Corolase PN-L and Corolase 7089 hydrolysates, except for those obtained under 300 MPa by the last protease. However, the immunochemical reaction was lower in the hydrolysis products obtained under HP than in those obtained at atmospheric pressure and after the HP treatment. On the contrary, no residual immunochemical reactivity associated with β-lactoglobulin was observed in the hydrolysates obtained by Alcalase and Neutrase under HP, and none was observed in any of the hydrolysis products obtained by Papain. The presence of traces of casein was not significant. These results suggest that HP combined with selected food-grade proteases is a treatment that can remove the antigenicity of whey protein hydrolysates for their use as ingredients of hypoallergenic infant formulae.

Document Type: Research Article

Affiliations: 1: Department of Science and Technology of Plant Food, Instituto del Frío, c/José Antonio Nováis 10, 28040-Madrid, España; Department of Science and Technology of Dairy Products, Instituto del Frío, c/José Antonio Nováis 10, 28040-Madrid, España 2: Department of Pharmacological Sciences, University of Milan, via Balzaretti 9, 20133 Milan, Italy 3: Department of Science and Technology of Plant Food, Instituto del Frío, c/José Antonio Nováis 10, 28040-Madrid, España 4: Department of Child and Maternal Medicine, The Macedonio Melloni Hospital, via Macedonio Melloni, 20129 Milan, Italy 5: Department of Science and Technology of Dairy Products, Instituto del Frío, c/José Antonio Nováis 10, 28040-Madrid, España

Publication date: July 1, 2006

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