Antigenic activity and epitope analysis of β-conglycinin hydrolyzed by pepsin

Soybean is among the 'big eight' allergenic foods, and β-conglycinin, the main antigenic protein of soybean, has high levels of antigenic activity. Why the antigenic activity of soybean β-conglycinin is not eliminated by enzymatic hydrolysis is not clear. In this study, changes in the molecular composition and antigenicity of β-conglycinin hydrolyzed by pepsin were analyzed and it was determined whether complete sequential epitopes exist in the resulting hydrolysates. The nature and antigenic activity of protein subunits obtained after β-conglycinin hydrolysis were also assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and competitive enzyme-linked immunosorbent assay, respectively.

The residual antigenic activity of β-conglycinin was 52%, α'- and α-subunits completely disappeared, the 49 kDa fraction partially disappeared, and peptides measuring 27 and 23 kDa were newly formed after 60 min of enzymatic hydrolysis. Prolonged enzymatic hydrolysis did not result in remarkable changes in these peptides; thus, the peptides show some resistance to enzymatic hydrolysis. The amino acid sequences of the peptide chains were analyzed by matrix-assisted laser desorption / ionization-time of flight mass spectrometry and aligned with the related sequences in the corresponding protein and antigen databases. Ten complete sequential epitopes were identified in the residual 49 kDa fraction, of these epitopes, two were from α-subunits and eight were from β-subunits.

The presence of complete sequential epitopes in hydrolysates obtained from the enzymatic hydrolysis of soybean is an important reason for the incomplete disappearance of the antigenic activity of β-conglycinin. © 2020 Society of Chemical Industry.