Enzymatic hydrolysis of buffalo casein enhances DPP-4 inhibition: Structural modifications and bioactive peptide identification

ABSTRACT: Dipeptidyl peptidase-4 (DPP-4), the enzyme responsible for the rapid degradation of incretin hormones, plays a pivotal role in blood glucose regulation, and its inhibition serves as an effective strategy for maintaining glucose homeostasis. The aim of this study was to investigate the effe...

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Main Authors: Ning An, Jing Yang, Yu Zhang, Huayi Suo, Jiajia Song
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:Journal of Dairy Science
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Online Access:http://www.sciencedirect.com/science/article/pii/S0022030224013341
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author Ning An
Jing Yang
Yu Zhang
Huayi Suo
Jiajia Song
author_facet Ning An
Jing Yang
Yu Zhang
Huayi Suo
Jiajia Song
author_sort Ning An
collection DOAJ
description ABSTRACT: Dipeptidyl peptidase-4 (DPP-4), the enzyme responsible for the rapid degradation of incretin hormones, plays a pivotal role in blood glucose regulation, and its inhibition serves as an effective strategy for maintaining glucose homeostasis. The aim of this study was to investigate the effect of enzymatic hydrolysis on the structure of buffalo casein and its DPP-4 inhibitory activity. Results demonstrated that Flavorzyme effectively hydrolyzed buffalo casein, as evidenced by scanning electron microscopy and electrophoretic analysis, with the degree of hydrolysis reaching its maximum value (20.05 ± 0.14%) after 3 h. The results of circular dichroism spectra, as well as endogenous and exogenous fluorescence spectra, indicated marked alterations in the secondary and tertiary structures of buffalo casein following enzymatic hydrolysis. Additionally, the DPP-4 inhibitory effect of buffalo casein was found to increase with longer hydrolysis times. The hydrolysate obtained after 3 h of hydrolysis demonstrated the highest level of inhibition, with a half-maximal inhibitory concentration (IC50) value of 1.04 mg/mL. The DPP-4 inhibitory peptide YPFPGPIPN, with an IC50 value of 0.88 mg/mL, was identified in the 1 to 3 kDa fraction of the 3-h hydrolysate. This peptide interacted with the active site of DPP-4 via hydrogen bonds, hydrophobic interactions, salt bridges, and π-cation interactions. This study offers a novel scientific foundation for the development of functional antidiabetic foods derived from buffalo casein.
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spelling doaj-art-833ee467e9634b809036cec77a96b9762025-08-20T02:01:01ZengElsevierJournal of Dairy Science0022-03022025-03-0110832169218110.3168/jds.2024-25552Enzymatic hydrolysis of buffalo casein enhances DPP-4 inhibition: Structural modifications and bioactive peptide identificationNing An0Jing Yang1Yu Zhang2Huayi Suo3Jiajia Song4College of Food Science, Southwest University, Chongqing 400715, ChinaSchool of Food Science and Engineering, Chongqing Technology and Business University, Chongqing 400067, ChinaCollege of Food Science, Southwest University, Chongqing 400715, ChinaCollege of Food Science, Southwest University, Chongqing 400715, ChinaCollege of Food Science, Southwest University, Chongqing 400715, China; Corresponding authorABSTRACT: Dipeptidyl peptidase-4 (DPP-4), the enzyme responsible for the rapid degradation of incretin hormones, plays a pivotal role in blood glucose regulation, and its inhibition serves as an effective strategy for maintaining glucose homeostasis. The aim of this study was to investigate the effect of enzymatic hydrolysis on the structure of buffalo casein and its DPP-4 inhibitory activity. Results demonstrated that Flavorzyme effectively hydrolyzed buffalo casein, as evidenced by scanning electron microscopy and electrophoretic analysis, with the degree of hydrolysis reaching its maximum value (20.05 ± 0.14%) after 3 h. The results of circular dichroism spectra, as well as endogenous and exogenous fluorescence spectra, indicated marked alterations in the secondary and tertiary structures of buffalo casein following enzymatic hydrolysis. Additionally, the DPP-4 inhibitory effect of buffalo casein was found to increase with longer hydrolysis times. The hydrolysate obtained after 3 h of hydrolysis demonstrated the highest level of inhibition, with a half-maximal inhibitory concentration (IC50) value of 1.04 mg/mL. The DPP-4 inhibitory peptide YPFPGPIPN, with an IC50 value of 0.88 mg/mL, was identified in the 1 to 3 kDa fraction of the 3-h hydrolysate. This peptide interacted with the active site of DPP-4 via hydrogen bonds, hydrophobic interactions, salt bridges, and π-cation interactions. This study offers a novel scientific foundation for the development of functional antidiabetic foods derived from buffalo casein.http://www.sciencedirect.com/science/article/pii/S0022030224013341enzymatic hydrolysisbuffalo caseindipeptidyl peptidase-4structure
spellingShingle Ning An
Jing Yang
Yu Zhang
Huayi Suo
Jiajia Song
Enzymatic hydrolysis of buffalo casein enhances DPP-4 inhibition: Structural modifications and bioactive peptide identification
Journal of Dairy Science
enzymatic hydrolysis
buffalo casein
dipeptidyl peptidase-4
structure
title Enzymatic hydrolysis of buffalo casein enhances DPP-4 inhibition: Structural modifications and bioactive peptide identification
title_full Enzymatic hydrolysis of buffalo casein enhances DPP-4 inhibition: Structural modifications and bioactive peptide identification
title_fullStr Enzymatic hydrolysis of buffalo casein enhances DPP-4 inhibition: Structural modifications and bioactive peptide identification
title_full_unstemmed Enzymatic hydrolysis of buffalo casein enhances DPP-4 inhibition: Structural modifications and bioactive peptide identification
title_short Enzymatic hydrolysis of buffalo casein enhances DPP-4 inhibition: Structural modifications and bioactive peptide identification
title_sort enzymatic hydrolysis of buffalo casein enhances dpp 4 inhibition structural modifications and bioactive peptide identification
topic enzymatic hydrolysis
buffalo casein
dipeptidyl peptidase-4
structure
url http://www.sciencedirect.com/science/article/pii/S0022030224013341
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AT yuzhang enzymatichydrolysisofbuffalocaseinenhancesdpp4inhibitionstructuralmodificationsandbioactivepeptideidentification
AT huayisuo enzymatichydrolysisofbuffalocaseinenhancesdpp4inhibitionstructuralmodificationsandbioactivepeptideidentification
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