Abstract
This study is the first to present an integrated approach involving in silico and in vitro protocols that was pursued to analyse an antioxidative potency of Gouda cheese with modified content of β-casein. Firstly, the predictions of the presence of antioxidant peptides in the casein sequences were computed using the BIOPEP-UWM database. Then, the antioxidative bioactivity of six variants of Gouda cheese (with reduced, normative, and increased content of β-casein at the initial and final stage of ripening) was assessed. Finally, the RP-HPLC-MS/MS was applied to identify antioxidative peptides in Gouda-derived water-soluble extracts (WSEs). Analyses were supported with the heatmaps and the computation of parameters describing the efficiency of proteolysis of caseins in the modified Gouda cheeses, i.e., the frequency and the relative frequency of the release of antioxidative fragments during cheese ripening (AEexp and Wexp., respectively). All Gouda cheese variants exhibited the antioxidative potential which differed depending on the assay employed. The highest antioxidative activity (ABTS·+ radical scavenging effect, FRAP, and Fe-chelating) was observed for WSEs derived from Gouda cheese with increased content of β-casein after the 60th day of ripening. The results obtained suggest the potential of Gouda cheese as the antioxidant-promoting food.