Immunoreactive properties of peptide fractions of cow whey milk proteins after enzymatic hydrolysis

• Published in: International journal of food science & technology Vol. 39; no. 8; pp. 839 - 850
• Main Authors: Wroblewska, B, Karamac, M, Amarowicz, R, Szymkiewicz, A, Troszynska, A, Kubicka, E
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01-10-2004; Blackwell Science
• Subjects: allergens; animal protein concentrates; beta-lactoglobulin; Biological and medical sciences; Enzymatic hydrolysis; enzymatic treatment; enzyme-linked immunosorbent assay; Food industries; Fundamental and applied biological sciences. Psychology; haptens; immune response; immunoassay; immunoreactivity; lactalbumin; lactoglobulins; milk allergy; Milk and cheese industries. Ice creams; papain; protein hydrolysates; proteinases; whey milk protein; whey protein; Hydrolysis; immunoreactivity; Milk protein; Peptides; Enzymatic hydrolysis; whey milk protein; Cow milk; Detection; Properties; Whey protein; Enzymatic reaction; immunoassay
• ISSN: 0950-5423; 1365-2621
• DOI: 10.1111/j.1365-2621.2004.00857.x

Commercial whey protein concentrate (WPC) was hydrolysed with either Alcalase 2.4 FG (Novo Nordisk), or papain (Sigma) (in one-step process) or with two enzymes (in two-step process) to determine the changes in the immunoreactivity of α-lactalbumin and β-lactoglobulin. Enzymatic hydrolysis of WPC was performed by pH-stat method. Hydrolysates were analysed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis, immunoblotting and size-exclusion chromatography (SE-HPLC). Immunoreactive properties of peptide fractions separated from the hydrolysates by fast protein liquid chromatography (FPLC) were determined using dot-immunobinding and enzyme-linked immunosorbent assay (ELISA) methods. Finally the sensory analysis was used to confirm organoleptic changes resulting from the application of different enzymes. The 'two-step' process was observed to be the most effective however allergenic epitopes were still present, as it was found by ELISA with anti-α-la and anti-β-lg antibodies. The addition of papain as the second enzyme in the hydrolysis process contributed to the improvement of the sensory properties of WPC hydrolysate as compared with the Alcalase hydrolysate. Alcalase-papain partially hydrolysated WPC can be found a promising base for production of the tolerogenic formula.