Effect of photo-oxidation of major milk proteins on protein structure and hydrolysis by chymosin

Changes in the protein structure of different major milk proteins in response to photo-oxidation and its effect on hydrolysis by chymosin were studied by determination of free amino-terminal groups and by peptide mapping using liquid chromatography coupled with mass spectrometry. Changes were seen in the formation of peptides by chymosin after photo-oxidation of all substrates studied, but the extent of changes varied between the different substrates. Oxidative changes involving tryptophan residues and formation of dityrosines were high in the casein, and conformational changes of both αS- and β-casein were indicated by changes in the micro-environment of the tryptophan residues after oxidation. A decreased accessibility of chymosin to oxidized caseins was reflected in a lower overall level of free amino-terminal groups, and changed peptide maps. Although β-lactoglobulin and lactoferrin showed marginal conformational changes, a higher level of free amino-terminal groups was observed, and a range of peptides were found to increase for these two proteins.