New Software for the Identification and Characterization of Peptides Generated during Fontina Cheese Ripening Using Mass Spectrometry Data

The aim of this work was to design and implement a new bioinformatics software which is able to identify the protein peptides from the peaks which arise from in-source or MS/MS fragmentation. The oligopeptide fraction was extracted from Fontina cheese at different ages of ripening and subsequently analyzed by LC/MS/MS. On the resulting total ion chromatograms, the peptides were identified by a method based both on the in-source fragmentation detectable with a single-quadrupole mass analyzer and by a new software which was developed. This software performs an in-silico digestion of the major milk proteins, it calculates all the possible peptide fragments which are generated by the loss of the first N- or C-terminal amino acids, and finally, it matches the experimental ion chromatogram with the in-silico which generated theoretical spectrum to identify the exact amino-acid protein sequence of the unknown oligopeptide. With this tool, the useful insights into the proteolytic processes which occur during Fontina cheese aging are obtained, which leads to a better knowledge about the functional features of the proteolysis end product.