Use of mass spectrometry to characterize proteolysis in cheese

A rapid method for characterizing proteolysis in different cheese varieties was developed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF-MS). pH 4.6-soluble extracts of different cheese varieties were sub-fractionated using ethanol and 70% ethanol-soluble extracts were analyzed using MALDI-ToF-MS. Sample analysis time was ∼3 min compared to ∼100 min for reversed-phase HPLC; moreover, the method yielded accurate molecular masses of peptides originating as a result of proteolysis. Small peptides (<3500 m/z) were monitored and data for relative intensities of peptides were analyzed using multidimensional scaling (MDS) to demonstrate the similarity between profiles obtained from different cheese varieties. The similarities between the profiles of MALDI-ToF mass spectra were demonstrated in a two-dimensional space by performing MDS on the similarity matrix. Use of MALDI-ToF-MS is thus a fast and effective method for monitoring small peptides produced in cheese as a result of proteolysis.