Proteolysis phenomena in cheese are a complex series of events leading to modifications in texture and flavor, both directly by the formation of peptides and free amino acids associated with development of desirable or undesirable taste and aroma, and indirectly by producing substrates for secondary biochemical reactions. The rate and extent of proteolysis is determined by several enzymatic activities coming from various sources, such as endogenous milk proteases, milk clotting enzymes, starter culture, contaminating microflora, etc., as well as technological food processing. Accordingly, dairy products can contain hundreds of different peptides at different levels of concentration. During the last years, several reviews and papers have been published on the analysis of intact milk proteins or peptides released from milk proteins in a wide variety of food products by Liquid Chromatography and Mass Spectrometry [1,2]. In this work, a novel bioinformatics approach coupled with nanoLC and electrospray ionization ion trap mass spectrometry is described for the identification of the oligopeptide fractions in Fior di Latte and cream cheese samples. The proposed method is based on scoring distribution associated to the protein hits at the top of each protein view report, originating from the database search of the complete LC-MS/MS run and followed by a refined identification by focusing on selected time-segments corresponding to the most intense peaks. This strategy allows the characterization of the most relevant peptides and minimizes the risk of false-positive identifications, reducing analysis time and costs, with the advantage of obtaining a lot of important information by a single LC-MS/MS run. [1] S. Sforza,V. Cavatorta, F. Lambertini, G. Galaverna, A. Dossena, R. Marchelli. J. Dairy Sci. 95 (2012) 3514–3526. [2] C. De Simone, G. Picariello, G. Mamone, P. Stiuso, A. Dicitore, D. Vanacore, L. Chianese, F. Addeo, P. Ferranti. J. Pept. Sci. 15 (2009) 251–258.
PEPTIDE PROFILING IN CHEESE SAMPLES BY LC‐TANDEM MASS SPECTROMETRY
NARDIELLO, DONATELLA;NATALE, ANNA;PALERMO, CARMEN;CONTE, AMALIA;LUCERA, ANNALISA;DEL NOBILE, MATTEO ALESSANDRO;CENTONZE, DIEGO
2013-01-01
Abstract
Proteolysis phenomena in cheese are a complex series of events leading to modifications in texture and flavor, both directly by the formation of peptides and free amino acids associated with development of desirable or undesirable taste and aroma, and indirectly by producing substrates for secondary biochemical reactions. The rate and extent of proteolysis is determined by several enzymatic activities coming from various sources, such as endogenous milk proteases, milk clotting enzymes, starter culture, contaminating microflora, etc., as well as technological food processing. Accordingly, dairy products can contain hundreds of different peptides at different levels of concentration. During the last years, several reviews and papers have been published on the analysis of intact milk proteins or peptides released from milk proteins in a wide variety of food products by Liquid Chromatography and Mass Spectrometry [1,2]. In this work, a novel bioinformatics approach coupled with nanoLC and electrospray ionization ion trap mass spectrometry is described for the identification of the oligopeptide fractions in Fior di Latte and cream cheese samples. The proposed method is based on scoring distribution associated to the protein hits at the top of each protein view report, originating from the database search of the complete LC-MS/MS run and followed by a refined identification by focusing on selected time-segments corresponding to the most intense peaks. This strategy allows the characterization of the most relevant peptides and minimizes the risk of false-positive identifications, reducing analysis time and costs, with the advantage of obtaining a lot of important information by a single LC-MS/MS run. [1] S. Sforza,V. Cavatorta, F. Lambertini, G. Galaverna, A. Dossena, R. Marchelli. J. Dairy Sci. 95 (2012) 3514–3526. [2] C. De Simone, G. Picariello, G. Mamone, P. Stiuso, A. Dicitore, D. Vanacore, L. Chianese, F. Addeo, P. Ferranti. J. Pept. Sci. 15 (2009) 251–258.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
