Characterizing lysinoalanine crosslinks in food systems: Discovery of a diagnostic ion in model peptides using MALDI mass spectrometry

Formation of lysinoalanine protein-protein crosslinks during food processing adversely impacts nutritional value. However, mapping lysinoalanine directly in food is challenging. We characterized the fragmentation pattern of lysinoalanine crosslinks in synthetic peptide models over a range of pH and time treatments using mass spectrometry. A putative diagnostic ion resulting from the cleavage of the & alpha;-carbon and & beta;-carbon of lysinoalanine is identified in MALDI MS/MS spectra. This represents the first step in mapping lysinoalanine in real food samples with higher precision than currently identifiable through standard or customized software. We then determined a correlated trend in the reduction of disulfide bonds and formation of lysinoalanine with increasing pH and time. Mapping lysinoalanine formation is critical to enhance our understanding of molecular processes impacting the nutritional value of foods, including notably in the development of protein alternatives that use alkaline treatment to extract protein isolates.

Automated summary

The formation of lysinoalanine protein-protein crosslinks during food processing negatively affects nutritional value. However, directly mapping lysinoalanine in food is challenging. This study characterized the fragmentation pattern of lysinoalanine crosslinks in synthetic peptide models under different pH and time treatments using mass spectrometry. A potential diagnostic ion resulting from the cleavage of the α-carbon and β-carbon of lysinoalanine was identified, providing a more precise method for mapping lysinoalanine in real food samples compared to current software.