Novel anti-oxidative peptides from equine hemoplasma protein hydrolysates: Purification, identification and protective effects on Caco-2 cells.
Abstract: In this study, we purified and identified antioxidant peptides from equine plasma protein hydrolysates and assessed their protective effects against HO-induced oxidative stress in Caco-2 cells. Four antioxidant peptides were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in equine plasma protein hydrolysate, namely: GTMVGC (567.69 Da), FGMTST (662.88 Da), VGYHSHF (847.01 Da) and ALSPFFKE (939.18 Da). Among them, ALSPFFKE showed the strongest antidigestive properties after modelled digestion studies. Moreover, ALSPFFKE enhanced intracellular superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) activities while significantly reducing reactive oxygen species accumulation and malondialdehyde formation in Caco-2 cells. The molecular docking analysis suggested that ALSPFFKE achieves regulation of the Keap1-Nrf2 pathway mainly by forming multiple hydrogen bonds and hydrophobic interactions with key amino acids (Arg380, Ser555, Gln530, Tyr334) in Keap1. These findings suggested that equine plasma peptides hold significant promise for the development of novel, potent, and stable antioxidant functional foods.
Copyright © 2025 Elsevier Ltd. All rights reserved.
Publication Date: 2025-02-05 PubMed ID: 39986787DOI: 10.1016/j.foodres.2025.115943Google Scholar: Lookup
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Summary
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This research article discusses the extraction and identification of antioxidant peptides from equine plasma protein hydrolysates, and their effectiveness in protecting against oxidative stress in Caco-2 cells.
Research Approach and Findings
- The researchers identified four antioxidant peptides from equine plasma protein hydrolysate by using liquid chromatography-tandem mass spectrometry (LC-MS/MS). These were GTMVGC, FGMTST, VGYHSHF and ALSPFFKE.
- Each peptide was evaluated on its properties and capabilities. The most promising was ALSPFFKE, which was found to have the strongest antidigestive characteristics after simulated digestion procedures.
Antioxidant Properties of Peptides
- During further studies, the peptide ALSPFFKE was seen to increase the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) within the cell. These enzymes are important for neutralising harmful reactive oxygen species.
- ALSPFFKE was also able to significantly reduce the formation of malondialdehyde and the accumulation of reactive oxygen species, both of which are indicators of oxidative stress.
Molecular Interaction
- The researchers conducted molecular docking analysis to investigate the mechanism of ALSPFFKE. The analysis suggests that ALSPFFKE might regulate the Keap1-Nrf2 pathway by forming hydrogen bonds and hydrophobic interactions with specific amino acids within Keap1.
- The Keap1-Nrf2 pathway is an important cellular defense mechanism against oxidative stress. Hence ALSPFFKE, through its interaction with this pathway, potentially contributes to its protective effect against oxidative stress.
Potential Applications
- Based on the findings of this study, the researchers propose that equine plasma peptides could be a promising avenue for creating novel antioxidant functional foods.
- Specifically, the peptide ALSPFFKE, with its powerful antioxidant properties and ability to survive digestion, could be used to create stable functional foods with potent antioxidant abilities.
Cite This Article
APA
Ma Z, Li Y, Zhao Z, Song Q, Wang Q, Lu S, Wang J.
(2025).
Novel anti-oxidative peptides from equine hemoplasma protein hydrolysates: Purification, identification and protective effects on Caco-2 cells.
Food Res Int, 204, 115943.
https://doi.org/10.1016/j.foodres.2025.115943 Publication
Researcher Affiliations
- Key Laboratory of Agricultural Product Processing and Quality Control of Specialty (Co-construction by Ministry and Province), School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China; Key Laboratory for Food Nutrition and Safety Control of Xinjiang Production and Construction Corps, School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China; Engineering Research Center of Storage and Processing of Xinjiang Characteristic Fruits and Vegetables, Ministry of Education, School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China.
- Key Laboratory of Agricultural Product Processing and Quality Control of Specialty (Co-construction by Ministry and Province), School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China; Engineering Research Center of Storage and Processing of Xinjiang Characteristic Fruits and Vegetables, Ministry of Education, School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China.
- Key Laboratory of Agricultural Product Processing and Quality Control of Specialty (Co-construction by Ministry and Province), School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China; Key Laboratory for Food Nutrition and Safety Control of Xinjiang Production and Construction Corps, School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China.
- Key Laboratory for Food Nutrition and Safety Control of Xinjiang Production and Construction Corps, School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China; Engineering Research Center of Storage and Processing of Xinjiang Characteristic Fruits and Vegetables, Ministry of Education, School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China.
- Key Laboratory of Agricultural Product Processing and Quality Control of Specialty (Co-construction by Ministry and Province), School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China; Key Laboratory for Food Nutrition and Safety Control of Xinjiang Production and Construction Corps, School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China; Engineering Research Center of Storage and Processing of Xinjiang Characteristic Fruits and Vegetables, Ministry of Education, School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China.
- Key Laboratory of Agricultural Product Processing and Quality Control of Specialty (Co-construction by Ministry and Province), School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China; Key Laboratory for Food Nutrition and Safety Control of Xinjiang Production and Construction Corps, School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China; Engineering Research Center of Storage and Processing of Xinjiang Characteristic Fruits and Vegetables, Ministry of Education, School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China.
- Key Laboratory of Agricultural Product Processing and Quality Control of Specialty (Co-construction by Ministry and Province), School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China; Key Laboratory for Food Nutrition and Safety Control of Xinjiang Production and Construction Corps, School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China; Engineering Research Center of Storage and Processing of Xinjiang Characteristic Fruits and Vegetables, Ministry of Education, School of Food Science and Technology, Shihezi University, Shihezi, Xinjiang 832000, China. Electronic address: 694817387@qq.com.
MeSH Terms
- Caco-2 Cells
- Humans
- Animals
- Antioxidants / pharmacology
- Antioxidants / isolation & purification
- Protein Hydrolysates / pharmacology
- Protein Hydrolysates / chemistry
- Horses
- Oxidative Stress / drug effects
- Peptides / isolation & purification
- Peptides / pharmacology
- Peptides / chemistry
- Molecular Docking Simulation
- Hydrogen Peroxide
- Reactive Oxygen Species / metabolism
- Glutathione Peroxidase / metabolism
- Tandem Mass Spectrometry
- Superoxide Dismutase / metabolism
- Catalase / metabolism
Conflict of Interest Statement
Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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