Direct nanoHPLC-ESI-QTOF MS/MS analysis of tryptic caseinophosphopeptides

Y. S. Zhu; R. J. FitzGerald

doi:10.1016/j.foodchem.2010.05.046

Direct nanoHPLC-ESI-QTOF MS/MS analysis of tryptic caseinophosphopeptides

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Abstract

Caseinophosphopeptides (CPPs) were generated following tryptic hydrolysis of sodium caseinate. Hydrolysate peptides were separated and identified using nano-HPLC ESI-QTOF MS/MS. Sequence coverage in the 3h hydrolysate was 79.4%, 55.6%, 80.9% and 68.1% for α_s1-, α_s2-, β- and κ-casein (CN), respectively. Variable levels of serine phosphorylation in β-CN f1-25 were observed in the 3h hydrolysate. Analysis of β-CN f1-25 4P demonstrated that this peptide was stable during the course of hydrolysis. The effect of heat treatment (75°C, 45min) at pH 6.0, 7.0 and 8.0 on the peptide profile of the 3h hydrolysate was studied. Compared to pH 6.0 and 8.0, least modification in phosphopeptide profiles was observed for the hydrolysate sample heated at pH 7.0. Different dephosphorylation and oxidation patterns were also observed following heat treatment at the three pH values. These results demonstrate that heat treatment, in addition to pH, has a major effect on both the phosphorylated and non-phosphorylated peptide profiles of CN hydrolysates.

Original language	English
Pages (from-to)	753-759
Number of pages	7
Journal	Food Chemistry
Volume	123
Issue number	3
DOIs	https://doi.org/10.1016/j.foodchem.2010.05.046
Publication status	Published - Dec 2010

Keywords

Caseinophosphopeptides
Heat treatment
Hydrolysis
Liquid chromatography
Mass spectrometry
PH effects

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10.1016/j.foodchem.2010.05.046

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title = "Direct nanoHPLC-ESI-QTOF MS/MS analysis of tryptic caseinophosphopeptides",

abstract = "Caseinophosphopeptides (CPPs) were generated following tryptic hydrolysis of sodium caseinate. Hydrolysate peptides were separated and identified using nano-HPLC ESI-QTOF MS/MS. Sequence coverage in the 3h hydrolysate was 79.4%, 55.6%, 80.9% and 68.1% for αs1-, αs2-, β- and κ-casein (CN), respectively. Variable levels of serine phosphorylation in β-CN f1-25 were observed in the 3h hydrolysate. Analysis of β-CN f1-25 4P demonstrated that this peptide was stable during the course of hydrolysis. The effect of heat treatment (75°C, 45min) at pH 6.0, 7.0 and 8.0 on the peptide profile of the 3h hydrolysate was studied. Compared to pH 6.0 and 8.0, least modification in phosphopeptide profiles was observed for the hydrolysate sample heated at pH 7.0. Different dephosphorylation and oxidation patterns were also observed following heat treatment at the three pH values. These results demonstrate that heat treatment, in addition to pH, has a major effect on both the phosphorylated and non-phosphorylated peptide profiles of CN hydrolysates.",

keywords = "Caseinophosphopeptides, Heat treatment, Hydrolysis, Liquid chromatography, Mass spectrometry, PH effects",

author = "Zhu, {Y. S.} and FitzGerald, {R. J.}",

year = "2010",

month = dec,

doi = "10.1016/j.foodchem.2010.05.046",

language = "English",

volume = "123",

pages = "753--759",

journal = "Food Chemistry",

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T1 - Direct nanoHPLC-ESI-QTOF MS/MS analysis of tryptic caseinophosphopeptides

AU - Zhu, Y. S.

AU - FitzGerald, R. J.

PY - 2010/12

Y1 - 2010/12

N2 - Caseinophosphopeptides (CPPs) were generated following tryptic hydrolysis of sodium caseinate. Hydrolysate peptides were separated and identified using nano-HPLC ESI-QTOF MS/MS. Sequence coverage in the 3h hydrolysate was 79.4%, 55.6%, 80.9% and 68.1% for αs1-, αs2-, β- and κ-casein (CN), respectively. Variable levels of serine phosphorylation in β-CN f1-25 were observed in the 3h hydrolysate. Analysis of β-CN f1-25 4P demonstrated that this peptide was stable during the course of hydrolysis. The effect of heat treatment (75°C, 45min) at pH 6.0, 7.0 and 8.0 on the peptide profile of the 3h hydrolysate was studied. Compared to pH 6.0 and 8.0, least modification in phosphopeptide profiles was observed for the hydrolysate sample heated at pH 7.0. Different dephosphorylation and oxidation patterns were also observed following heat treatment at the three pH values. These results demonstrate that heat treatment, in addition to pH, has a major effect on both the phosphorylated and non-phosphorylated peptide profiles of CN hydrolysates.

AB - Caseinophosphopeptides (CPPs) were generated following tryptic hydrolysis of sodium caseinate. Hydrolysate peptides were separated and identified using nano-HPLC ESI-QTOF MS/MS. Sequence coverage in the 3h hydrolysate was 79.4%, 55.6%, 80.9% and 68.1% for αs1-, αs2-, β- and κ-casein (CN), respectively. Variable levels of serine phosphorylation in β-CN f1-25 were observed in the 3h hydrolysate. Analysis of β-CN f1-25 4P demonstrated that this peptide was stable during the course of hydrolysis. The effect of heat treatment (75°C, 45min) at pH 6.0, 7.0 and 8.0 on the peptide profile of the 3h hydrolysate was studied. Compared to pH 6.0 and 8.0, least modification in phosphopeptide profiles was observed for the hydrolysate sample heated at pH 7.0. Different dephosphorylation and oxidation patterns were also observed following heat treatment at the three pH values. These results demonstrate that heat treatment, in addition to pH, has a major effect on both the phosphorylated and non-phosphorylated peptide profiles of CN hydrolysates.

KW - Caseinophosphopeptides

KW - Heat treatment

KW - Hydrolysis

KW - Liquid chromatography

KW - Mass spectrometry

KW - PH effects

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DO - 10.1016/j.foodchem.2010.05.046

M3 - Article

AN - SCOPUS:77954621834

SN - 0308-8146

VL - 123

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JO - Food Chemistry

JF - Food Chemistry

IS - 3

ER -

Direct nanoHPLC-ESI-QTOF MS/MS analysis of tryptic caseinophosphopeptides

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Keywords

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