TY - JOUR
T1 - Identification of short peptide sequences in the nanofiltration permeate of a bioactive whey protein hydrolysate
AU - Le Maux, Solène
AU - Nongonierma, Alice B.
AU - Murray, Brian
AU - Kelly, Phil M.
AU - FitzGerald, Richard J.
N1 - Publisher Copyright:
© 2015 Elsevier Ltd.
PY - 2015/11/1
Y1 - 2015/11/1
N2 - Short peptides in food protein hydrolysates are of significant interest as they may be highly bioactive whilst also being bioavailable. A dipeptidyl peptidase IV (DPP-IV) inhibitory whey protein hydrolysate (WPH) was fractionated using nanofiltration (NF) with a 200Da MWCO membrane. The DPP-IV half maximal inhibitory concentration of the NF permeate (IC50=0.66±0.08mgprotein equivalentmL-1) was significantly more potent (P>0.05) than that of the starting WPH (IC50=0.94±0.24mgprotein equivalentmL-1) and associated retentate (IC50=0.82±0.13mgprotein equivalentmL-1). This confirmed the contribution of short peptides within the NF permeate to the overall DPP-IV inhibitory activity. An hydrophilic interaction liquid chromatography (HILIC-) and reverse-phase (RP-) liquid chromatography tandem mass spectrometry (LC-MS/MS) strategy, based on two retention time models, allowed detection of eight free amino acids and eight di- to tetrapeptides in the NF permeate. The potential sequences of the peptides within the NF permeate were then ranked on the basis of their highest probability of occurrence. A confirmatory study with synthetic peptides showed that valine-alanine (VA), valine-leucine (VL), tryptophan-leucine (WL) and tryptophan-isoleucine (WI) displayed DPP-IV IC50 values <170μM. The NF and LC-MS strategies employed herein represent a new approach for the targeted identification of short peptides within bioactive food protein hydrolysates.
AB - Short peptides in food protein hydrolysates are of significant interest as they may be highly bioactive whilst also being bioavailable. A dipeptidyl peptidase IV (DPP-IV) inhibitory whey protein hydrolysate (WPH) was fractionated using nanofiltration (NF) with a 200Da MWCO membrane. The DPP-IV half maximal inhibitory concentration of the NF permeate (IC50=0.66±0.08mgprotein equivalentmL-1) was significantly more potent (P>0.05) than that of the starting WPH (IC50=0.94±0.24mgprotein equivalentmL-1) and associated retentate (IC50=0.82±0.13mgprotein equivalentmL-1). This confirmed the contribution of short peptides within the NF permeate to the overall DPP-IV inhibitory activity. An hydrophilic interaction liquid chromatography (HILIC-) and reverse-phase (RP-) liquid chromatography tandem mass spectrometry (LC-MS/MS) strategy, based on two retention time models, allowed detection of eight free amino acids and eight di- to tetrapeptides in the NF permeate. The potential sequences of the peptides within the NF permeate were then ranked on the basis of their highest probability of occurrence. A confirmatory study with synthetic peptides showed that valine-alanine (VA), valine-leucine (VL), tryptophan-leucine (WL) and tryptophan-isoleucine (WI) displayed DPP-IV IC50 values <170μM. The NF and LC-MS strategies employed herein represent a new approach for the targeted identification of short peptides within bioactive food protein hydrolysates.
KW - Bioactive peptides
KW - Dipeptidyl peptidase IV inhibition
KW - Mass spectrometry
KW - Retention time
KW - Short peptides
UR - http://www.scopus.com/inward/record.url?scp=84949088240&partnerID=8YFLogxK
U2 - 10.1016/j.foodres.2015.09.012
DO - 10.1016/j.foodres.2015.09.012
M3 - Article
AN - SCOPUS:84949088240
SN - 0963-9969
VL - 77
SP - 534
EP - 539
JO - Food Research International
JF - Food Research International
ER -