Regulation of action sites for reducing the allergenicity of pea protein based on enzymatic hydrolysis with Alcalase

Enzymatic hydrolysis could reduce the allergenicity of pea protein. Box-Behnken model was used to extract vicilin with the lowest and highest allergenicity, and enzymatic hydrolysis, electrophoresis, spectroscopy, bioinformatics, and peptidomics of Nano-LC-MS/MS were utilized to explore the relationship between reduced allergenicity and structural changes. After enzymatic hydrolysis, the allergenicity of L-vicilin hydrolysates (L-VHs) and H-vicilin hydrolysates (H-VHs) decreased significantly (P < 0.05). Furthermore, large-molecular-weight subunits in L-vicilin and H-vicilin were decomposed into <11 kDa peptides, and their surface hydrophobicity were increased. The OH, NH, and CO groups underwent stretching vibrations, and α-helix and β-sheet were transformed into β-turn and random coils. Additionally, linear epitopes of P13918, D3VND7, D3VNE2, and P02856 in L-VHs and H-VHs were cut into different fragments. Among them, distinct linear epitope fragments of them might be responsible for the difference in their allergenicity. Therefore, enzymatic hydrolysis with Alclase could effectively reduce the allergenicity of vicilin by regulating sensitization sites.