Isolation of oligopeptides from the water-soluble extract of goat cheese and their identification by mass spectrometry.
A procedure for the separation and identification of small peptides from the water-soluble fraction of a goat cheese was developed. The water-soluble extract was ultrafiltered (1000 Da membrane cutoff), and peptides were isolated by sequential chromatography: size exclusion chromatography (HPLC-grade water), anion exchange chromatography (phosphate buffer gradient), and semipreparative reverse-phase high-performance liquid chromatography (water/acetonitrile gradient). The fractions obtained were analyzed by combined mass spectrometry methods including electrospray ionization, liquid secondary ionization, and tandem mass spectrometry to identify and to confirm the sequences of 28 tri- to octapeptides naturally appearing in goat cheese during ripening. Among these peptides, 26 are produced by degradation of caseins but do not correspond to the known specific cleavages due to chymosin. Only low correlation was found between hydrophobicity of peptides and HPLC elution time with acetonitrile gradient.
Sommerer
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N
INRA Laboratoire de Recherches Sur les Arômes, 17 rue Sully, F-21065 Dijon Cedex, France.
Salles
C
C
Promé
D
D
Promé
J C
JC
Le Quéré
J L
JL
D000595:Amino Acid Sequence; D000818:Animals; D002611:Cheese / Q000032:analysis*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D006041:Goats; D013058:Mass Spectrometry; D009842:Oligopeptides / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D012995:Solubility; D014867:Water
D009842:Oligopeptides; D014867:Water