Hemoglobin-Derived Antioxidant Peptides from Beef Cattle in Ningxia: Identification Based on Computerized Screening and Molecular Docking
In this study, three enzymatic hydrolysis methods, alkaline protease (E1), pepsin + trypsin (E2), and alkaline protease + pepsin + trypsin (E3), were used to prepare hemoglobin hydrolysates from Ningxia beef cattle, which were screened for their antioxidant activity. Amino acid analysis and liquid c...
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China Food Publishing Company
2025-01-01
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| author | XU Feiran, WANG Yu, WANG Xiaojing, CUI Wei, WANG Yongkun, ZHOU Hui, WANG Zhaoming, CAI Kezhou, ZHANG Bao, XU Baocai |
| author_facet | XU Feiran, WANG Yu, WANG Xiaojing, CUI Wei, WANG Yongkun, ZHOU Hui, WANG Zhaoming, CAI Kezhou, ZHANG Bao, XU Baocai |
| author_sort | XU Feiran, WANG Yu, WANG Xiaojing, CUI Wei, WANG Yongkun, ZHOU Hui, WANG Zhaoming, CAI Kezhou, ZHANG Bao, XU Baocai |
| collection | DOAJ |
| description | In this study, three enzymatic hydrolysis methods, alkaline protease (E1), pepsin + trypsin (E2), and alkaline protease + pepsin + trypsin (E3), were used to prepare hemoglobin hydrolysates from Ningxia beef cattle, which were screened for their antioxidant activity. Amino acid analysis and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used to identify the amino acid compositions and sequences of the hydrolysates with molecular mass < 3 kDa. The potential antioxidant peptides were selected by computerized analysis and their activities were verified. Their action mechanism was studied by molecular simulation. The results showed that E3 exhibited the highest hydrolysis efficiency, and the < 3 kDa fraction obtained from the ultrafiltration of the resulting hydrolysate showed better antioxidant activity with half maximal inhibitory concentration (IC50) of (18.382 ± 0.023) mg/mL and (0.246 ± 0.012) mg/mL against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) cationic radical, respectively. The < 3 kDa fraction was rich in hydrophobic and aromatic amino acids, and four peptides with the best antioxidant activity were selected and identified, whose amino acid sequences were DWGHSW, VFYPW, GTGW and TYFPHF, respectively. Among them, VFYPW showed the highest DPPH radical and ABTS cationic radical scavenging capacity with IC50 of (0.350 ± 0.013) mg/mL and (9.482 ± 0.122) µg/mL, followed by DWGHSW. DWGHSW and VFYPW were found to form stable complexes with Keap1 protein (PDB ID: 2FLU) through hydrogen bonds and hydrophobic interactions. In summary, this study provides a theoretical basis for the application of hemoglobin peptides from Ningxia beef cattle as natural antioxidant supplements and contributes to the high-value utilization of blood resources from Ningxia beef cattle. |
| format | Article |
| id | doaj-art-c50355a495c74d858961b172f9a698c1 |
| institution | Kabale University |
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| language | English |
| publishDate | 2025-01-01 |
| publisher | China Food Publishing Company |
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| spelling | doaj-art-c50355a495c74d858961b172f9a698c12025-02-05T09:08:22ZengChina Food Publishing CompanyShipin Kexue1002-66302025-01-0146210811710.7506/spkx1002-6630-20240430-295Hemoglobin-Derived Antioxidant Peptides from Beef Cattle in Ningxia: Identification Based on Computerized Screening and Molecular DockingXU Feiran, WANG Yu, WANG Xiaojing, CUI Wei, WANG Yongkun, ZHOU Hui, WANG Zhaoming, CAI Kezhou, ZHANG Bao, XU Baocai0(1. Key Laboratory for Animal Food Green Manufacturing and Resource Mining of Anhui Province, School of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, China; 2. Ningxia Institute of Quality Standards and Testing Technology for Agricultural product, Yinchuan 750002, China)In this study, three enzymatic hydrolysis methods, alkaline protease (E1), pepsin + trypsin (E2), and alkaline protease + pepsin + trypsin (E3), were used to prepare hemoglobin hydrolysates from Ningxia beef cattle, which were screened for their antioxidant activity. Amino acid analysis and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used to identify the amino acid compositions and sequences of the hydrolysates with molecular mass < 3 kDa. The potential antioxidant peptides were selected by computerized analysis and their activities were verified. Their action mechanism was studied by molecular simulation. The results showed that E3 exhibited the highest hydrolysis efficiency, and the < 3 kDa fraction obtained from the ultrafiltration of the resulting hydrolysate showed better antioxidant activity with half maximal inhibitory concentration (IC50) of (18.382 ± 0.023) mg/mL and (0.246 ± 0.012) mg/mL against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) cationic radical, respectively. The < 3 kDa fraction was rich in hydrophobic and aromatic amino acids, and four peptides with the best antioxidant activity were selected and identified, whose amino acid sequences were DWGHSW, VFYPW, GTGW and TYFPHF, respectively. Among them, VFYPW showed the highest DPPH radical and ABTS cationic radical scavenging capacity with IC50 of (0.350 ± 0.013) mg/mL and (9.482 ± 0.122) µg/mL, followed by DWGHSW. DWGHSW and VFYPW were found to form stable complexes with Keap1 protein (PDB ID: 2FLU) through hydrogen bonds and hydrophobic interactions. In summary, this study provides a theoretical basis for the application of hemoglobin peptides from Ningxia beef cattle as natural antioxidant supplements and contributes to the high-value utilization of blood resources from Ningxia beef cattle.https://www.spkx.net.cn/fileup/1002-6630/PDF/2025-46-2-013.pdfcomputerized screening; molecular simulation; hemoglobin of ningxia beef cattle; antioxidant peptides; identification |
| spellingShingle | XU Feiran, WANG Yu, WANG Xiaojing, CUI Wei, WANG Yongkun, ZHOU Hui, WANG Zhaoming, CAI Kezhou, ZHANG Bao, XU Baocai Hemoglobin-Derived Antioxidant Peptides from Beef Cattle in Ningxia: Identification Based on Computerized Screening and Molecular Docking Shipin Kexue computerized screening; molecular simulation; hemoglobin of ningxia beef cattle; antioxidant peptides; identification |
| title | Hemoglobin-Derived Antioxidant Peptides from Beef Cattle in Ningxia: Identification Based on Computerized Screening and Molecular Docking |
| title_full | Hemoglobin-Derived Antioxidant Peptides from Beef Cattle in Ningxia: Identification Based on Computerized Screening and Molecular Docking |
| title_fullStr | Hemoglobin-Derived Antioxidant Peptides from Beef Cattle in Ningxia: Identification Based on Computerized Screening and Molecular Docking |
| title_full_unstemmed | Hemoglobin-Derived Antioxidant Peptides from Beef Cattle in Ningxia: Identification Based on Computerized Screening and Molecular Docking |
| title_short | Hemoglobin-Derived Antioxidant Peptides from Beef Cattle in Ningxia: Identification Based on Computerized Screening and Molecular Docking |
| title_sort | hemoglobin derived antioxidant peptides from beef cattle in ningxia identification based on computerized screening and molecular docking |
| topic | computerized screening; molecular simulation; hemoglobin of ningxia beef cattle; antioxidant peptides; identification |
| url | https://www.spkx.net.cn/fileup/1002-6630/PDF/2025-46-2-013.pdf |
| work_keys_str_mv | AT xufeiranwangyuwangxiaojingcuiweiwangyongkunzhouhuiwangzhaomingcaikezhouzhangbaoxubaocai hemoglobinderivedantioxidantpeptidesfrombeefcattleinningxiaidentificationbasedoncomputerizedscreeningandmoleculardocking |