Purification, identification, and in silico screening of a multifunctional octapeptide from semen armeniacae glutelin-2 hydrolysates: restraining mechanisms to Keap1 and ACE, stability, and ferrous-transport efficiency

IntroductionSemen armeniacae is a traditional homologous material of medicine and food, but data on its multifunctional peptides are little.MethodsIn this study, semen armeniacae glutelin-2 was hydrolyzed by alcalase and trypsin assisted with ultrasound. Antihypertensive and antioxidant peptides wit...

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Main Authors: Ziqing Jin, Ling Dang, Yan Li, Chen Feng, Xinling Song, Zhihui Wei, Jie Liu, Hao Wang, Yichan Zhang
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-04-01
Series:Frontiers in Nutrition
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Online Access:https://www.frontiersin.org/articles/10.3389/fnut.2025.1571161/full
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author Ziqing Jin
Ling Dang
Yan Li
Chen Feng
Xinling Song
Zhihui Wei
Jie Liu
Hao Wang
Yichan Zhang
author_facet Ziqing Jin
Ling Dang
Yan Li
Chen Feng
Xinling Song
Zhihui Wei
Jie Liu
Hao Wang
Yichan Zhang
author_sort Ziqing Jin
collection DOAJ
description IntroductionSemen armeniacae is a traditional homologous material of medicine and food, but data on its multifunctional peptides are little.MethodsIn this study, semen armeniacae glutelin-2 was hydrolyzed by alcalase and trypsin assisted with ultrasound. Antihypertensive and antioxidant peptides with ferrous-binding activity were isolated, identified, and in silico screened from the hydrolysates, and the action mechanisms against Keap1 and angiotensin-I-converting enzyme (ACE), gastrointestinal stability, and ferrous-binding capacity were studied.Results and discussionAfter Sephadex G-15 isolation, electrospray ionization mass spectrometry, and AHTpin and Peptide Ranker database screening, a safe multifunctional octapeptide: Pro-Val-Asp-Phe-Ala-Gly-Phe-Tyr (PVDFAGFY), was obtained. The capacities of PVDFAGFY to restrain ACE, chelate ferrous ions, and quench hydroxyl radical were IC50:105.61 μmol/L, 11.67 mg/g, and 97.67%, respectively. PVDFAGFY restrained ACE via competitively linking to its catalytic (His383) and/or crucial binding sites (Gln281, Lys511, Tyr523, Tyr520, or Ala354), and it can inhibit the Keap1-Nrf2 interaction by binding to 6 residues of Keap1. Ferrous ions were primarily chelated by γ-hydroxyl, carboxyl, and/or amino groups of PVDFAGFY via ionic forces. Gastrointestinal hydrolysis did not decrease the capacity of PVDFAGFY to antioxidant and restrain ACE (p > 0.05). The ACE inhibition model and activity of PVDFAGFY were not altered by iron chelation; however, PVDFAGFY-ferrous chelate showed lower hydroxyl and ABTS radical quenching capacity and ferric reducing ability than PVDFAGFY (p < 0.05). The gastrointestinal stability and transmembrane absorption of ferrous ions were increased by PVDFAGFY (p < 0.05). Thus, PVDFAGFY may be exploited as ingredients of hypotensive, antioxidant, and/or iron supplementary agents, but in vivo antioxidant and hypotensive efficiencies need further study.
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spelling doaj-art-6decb8ee882747b09f6ac899d8c7fd292025-08-20T01:51:55ZengFrontiers Media S.A.Frontiers in Nutrition2296-861X2025-04-011210.3389/fnut.2025.15711611571161Purification, identification, and in silico screening of a multifunctional octapeptide from semen armeniacae glutelin-2 hydrolysates: restraining mechanisms to Keap1 and ACE, stability, and ferrous-transport efficiencyZiqing Jin0Ling Dang1Yan Li2Chen Feng3Xinling Song4Zhihui Wei5Jie Liu6Hao Wang7Yichan Zhang8Food Science College of Shanxi Normal University, Taiyuan, ChinaShanxi Technology and Business University, Taiyuan, ChinaFood Science College of Shanxi Normal University, Taiyuan, ChinaFood Science College of Shanxi Normal University, Taiyuan, ChinaFood Science College of Shanxi Normal University, Taiyuan, ChinaFood Science College of Shanxi Normal University, Taiyuan, ChinaFood Science College of Shanxi Normal University, Taiyuan, ChinaFood Science College of Shanxi Normal University, Taiyuan, ChinaFood Science College of Shanxi Normal University, Taiyuan, ChinaIntroductionSemen armeniacae is a traditional homologous material of medicine and food, but data on its multifunctional peptides are little.MethodsIn this study, semen armeniacae glutelin-2 was hydrolyzed by alcalase and trypsin assisted with ultrasound. Antihypertensive and antioxidant peptides with ferrous-binding activity were isolated, identified, and in silico screened from the hydrolysates, and the action mechanisms against Keap1 and angiotensin-I-converting enzyme (ACE), gastrointestinal stability, and ferrous-binding capacity were studied.Results and discussionAfter Sephadex G-15 isolation, electrospray ionization mass spectrometry, and AHTpin and Peptide Ranker database screening, a safe multifunctional octapeptide: Pro-Val-Asp-Phe-Ala-Gly-Phe-Tyr (PVDFAGFY), was obtained. The capacities of PVDFAGFY to restrain ACE, chelate ferrous ions, and quench hydroxyl radical were IC50:105.61 μmol/L, 11.67 mg/g, and 97.67%, respectively. PVDFAGFY restrained ACE via competitively linking to its catalytic (His383) and/or crucial binding sites (Gln281, Lys511, Tyr523, Tyr520, or Ala354), and it can inhibit the Keap1-Nrf2 interaction by binding to 6 residues of Keap1. Ferrous ions were primarily chelated by γ-hydroxyl, carboxyl, and/or amino groups of PVDFAGFY via ionic forces. Gastrointestinal hydrolysis did not decrease the capacity of PVDFAGFY to antioxidant and restrain ACE (p > 0.05). The ACE inhibition model and activity of PVDFAGFY were not altered by iron chelation; however, PVDFAGFY-ferrous chelate showed lower hydroxyl and ABTS radical quenching capacity and ferric reducing ability than PVDFAGFY (p < 0.05). The gastrointestinal stability and transmembrane absorption of ferrous ions were increased by PVDFAGFY (p < 0.05). Thus, PVDFAGFY may be exploited as ingredients of hypotensive, antioxidant, and/or iron supplementary agents, but in vivo antioxidant and hypotensive efficiencies need further study.https://www.frontiersin.org/articles/10.3389/fnut.2025.1571161/fullsemen armeniacae glutelin-2 octapeptidedual enzymolysisangiotensin-I-converting enzymeKeap1inhibition mechanismsantioxidant
spellingShingle Ziqing Jin
Ling Dang
Yan Li
Chen Feng
Xinling Song
Zhihui Wei
Jie Liu
Hao Wang
Yichan Zhang
Purification, identification, and in silico screening of a multifunctional octapeptide from semen armeniacae glutelin-2 hydrolysates: restraining mechanisms to Keap1 and ACE, stability, and ferrous-transport efficiency
Frontiers in Nutrition
semen armeniacae glutelin-2 octapeptide
dual enzymolysis
angiotensin-I-converting enzyme
Keap1
inhibition mechanisms
antioxidant
title Purification, identification, and in silico screening of a multifunctional octapeptide from semen armeniacae glutelin-2 hydrolysates: restraining mechanisms to Keap1 and ACE, stability, and ferrous-transport efficiency
title_full Purification, identification, and in silico screening of a multifunctional octapeptide from semen armeniacae glutelin-2 hydrolysates: restraining mechanisms to Keap1 and ACE, stability, and ferrous-transport efficiency
title_fullStr Purification, identification, and in silico screening of a multifunctional octapeptide from semen armeniacae glutelin-2 hydrolysates: restraining mechanisms to Keap1 and ACE, stability, and ferrous-transport efficiency
title_full_unstemmed Purification, identification, and in silico screening of a multifunctional octapeptide from semen armeniacae glutelin-2 hydrolysates: restraining mechanisms to Keap1 and ACE, stability, and ferrous-transport efficiency
title_short Purification, identification, and in silico screening of a multifunctional octapeptide from semen armeniacae glutelin-2 hydrolysates: restraining mechanisms to Keap1 and ACE, stability, and ferrous-transport efficiency
title_sort purification identification and in silico screening of a multifunctional octapeptide from semen armeniacae glutelin 2 hydrolysates restraining mechanisms to keap1 and ace stability and ferrous transport efficiency
topic semen armeniacae glutelin-2 octapeptide
dual enzymolysis
angiotensin-I-converting enzyme
Keap1
inhibition mechanisms
antioxidant
url https://www.frontiersin.org/articles/10.3389/fnut.2025.1571161/full
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