Optimizing β-Lactoglobulin antigenicity through single enzyme hydrolysis: Exploring structural changes and effects on linear epitopes

β-lactoglobulin (β-LG) is the major allergen in dairy products, but research on the optimal conditions for antigen reduction in β-LG using different enzymes remains limited. Therefore, this study aims to investigate the antigenicity, structural characteristics, and peptide distribution of advantageous protease hydrolysates capable of eliminating the allergenic epitopes of β-LG selected via bioinformatics tools. The results showed that under optimal enzymatic hydrolysis conditions, the antigen reduction rates for the four advantageous proteases acting on β-LG were 47.37 % (pepsin), 33.54 % (chymotrypsin A), 38.71 % (papain), and 45.91 % (stem bromelain), respectively. The four proteases effectively degraded β-LG, causing shorter peptide chain formation, reduced content of highly ordered α-helix, decreased fluorescence intensity, and lower surface hydrophobicity. Furthermore, they cleaved the linear epitopes of β-LG into peptides of varying sizes, leading to different antigen reduction rates among the hydrolysates. These findings provide a theoretical basis for developing targeted enzymatic hydrolysis technologies and low-allergenicity dairy-based materials. [Display omitted] •Screen out the theoretically advantageous enzymes.•Optimise hydrolysis process for the highest antigen reduction rate of β-LG.•Enzymatic hydrolysis degraded β-LG and changed the structures.•Enzymatic hydrolysis disrupted the antigenic epitopes of β-LG.•Use peptidomics to analyse residual antigenic epitopes.