Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)The effect of combined treatments of hydrolysis with different proteases, and subsequent polymerization with transglutaminase on the antigenic activity of beta-Lg was studied. For the hydrolysis of beta-Lg using Alcalase, Neutrase or bromelain, the reaction conditions were 3 % beta-Lg and enzyme:substrate 25 U g(-1) of protein, as was defined using factorial study. Under these conditions, the degree of hydrolysis (DH) of the hydrolysates was 12.6 % when obtained with Alcalase and approximately 4 % with Neutrase or bromelain. Post-hydrolysis polymerization did not result in an increase in molecular mass of the protein, but these samples presented a lower DH, determined by trinitrobenzenosulfonic acid (TNBS) method, suggesting that polymerization had occurred. Hydrolysis with the three enzymes reduced the beta-Lg antigenicity, as evaluated by ELISA and immunoblotting analyses. The IgE-binding responses were practically null (< 9 mu g mL(-1)), 22.82 and 55.73 mu g mL(-1) towards the hydrolysates obtained with Alcalase, bromelain, and Neutrase, respectively. The post-hydrolysis polymerization increased or had no significant effect (P a parts per thousand yen 0.05) on the antigenic response of the hydrolysates.2355801809Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)