Analysis of the endogenous peptide profile of milk: identification of 248 mainly casein-derived peptides.
Milk is an excellent source of bioactive peptides. However, the composition of the native milk peptidome has only been partially elucidated. The present study applied matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) directly or after prefractionation of the milk peptides by reverse-phase high-performance liquid chromatography (RP-HPLC) or OFFGEL fractionation for the comprehensive analysis of the peptide profile of raw milk. The peptide sequences were determined by MALDI-TOF/TOF or nano-ultra-performance liquid chromatography-nanoelectrospray ionization-LTQ-Orbitrap-MS. Direct MALDI-TOF-MS analysis led to the assignment of 57 peptides. Prefractionation by both complementary methods led to the assignment of another 191 peptides. Most peptides originate from α(S1)-casein, followed by β-casein, and α(S2)-casein. κ-Casein and whey proteins seem to play only a minor role as peptide precursors. The formation of many, but not all, peptides could be explained by the activity of the endogenous peptidases, plasmin or cathepsin D, B, and G. Database searches revealed the presence of 22 peptides with established physiological function, including those with angiotensin-converting-enzyme (ACE) inhibitory, immunomodulating, or antimicrobial activity.
Department of Chemistry and Pharmacy, Food Chemistry, Emil Fischer Center, University of Erlangen-Nuremberg , Schuhstrasse 19, 91052 Erlangen, Germany.
Florian
F
Baum
Maria
M
Fedorova
Jennifer
J
Ebner
Ralf
R
Hoffmann
Monika
M
Pischetsrieder
D000595:Amino Acid Sequence; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D000818:Animals; D000890:Anti-Infective Agents / Q000737:chemistry / Q000302:isolation & purification*; D002364:Caseins / Q000737:chemistry / Q000302:isolation & purification*; D005591:Chemical Fractionation; D056148:Chromatography, Reverse-Phase; D004579:Electron Transport; D004583:Electrons; D005341:Fibrinolysin / Q000737:chemistry / Q000302:isolation & purification; D007155:Immunologic Factors / Q000737:chemistry / Q000302:isolation & purification*; D008892:Milk / Q000737:chemistry*; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000806:Angiotensin-Converting Enzyme Inhibitors; D000890:Anti-Infective Agents; D002364:Caseins; D007155:Immunologic Factors; D010455:Peptides; D005341:Fibrinolysin