基于计算机筛选与分子模拟技术的宁夏肉牛血红蛋白源抗氧化肽的鉴定

doi:10.7506/spkx1002-6630-20240430-295

食品科学 ›› 2025, Vol. 46 ›› Issue (2): 108-117.doi: 10.7506/spkx1002-6630-20240430-295

基于计算机筛选与分子模拟技术的宁夏肉牛血红蛋白源抗氧化肽的鉴定

徐斐然，王玉，王晓静，崔伟，王永坤，周辉，王兆明，蔡克周，章宝，徐宝才

（1.合肥工业大学食品与生物工程学院，动物源食品绿色制造与资源挖掘安徽省重点实验室，安徽合肥 230601；2.宁夏农产品质量标准与检测技术研究所，宁夏银川 750002）

出版日期:2025-01-25 发布日期:2024-12-30
基金资助:
宁夏回族自治区重点研发计划项目（2023BDE03012）；2023年度宁夏农业科技自主创新专项（NKYG-23-04）； “十四五”国家重点研发计划重点专项（2021YFD2100804）

Hemoglobin-Derived Antioxidant Peptides from Beef Cattle in Ningxia: Identification Based on Computerized Screening and Molecular Docking

XU Feiran, WANG Yu, WANG Xiaojing, CUI Wei, WANG Yongkun, ZHOU Hui, WANG Zhaoming, CAI Kezhou, ZHANG Bao, XU Baocai

(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)

Online:2025-01-25 Published:2024-12-30

摘要/Abstract

摘要： 采用3 种酶解法（碱性蛋白酶（E1组）、胃蛋白酶＋胰蛋白酶（E2组）、碱性蛋白酶＋胃蛋白酶＋胰蛋白酶（E3组））制备宁夏肉牛血红蛋白酶解产物，并筛选出抗氧化活性最佳组分；利用氨基酸分析和液相色谱-串联质谱鉴定分子质量＜3 kDa酶解产物的氨基酸组成和序列，利用计算机分析技术筛选出潜在的抗氧化肽，并验证其活性；通过分子模拟技术进一步研究宁夏肉牛血红蛋白抗氧化肽的作用机理。结果表明，E3组酶解效果最佳，超滤后＜3 kDa的组分表现出更佳的抗氧化活性（1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl，DPPH）自由基和2,2’-联氮-双(3-乙基苯并噻唑啉-6-磺酸)（2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)，ABTS）阳离子自由基清除率的半数抑制浓度（half maximal inhibitory concentration，IC50）分别为（18.382±0.023）mg/mL和（0.246±0.012）mg/mL）；E3（分子质量＜3 kDa）组富含疏水性和芳香族氨基酸，且鉴定和筛选出4 条活性最佳的抗氧化肽，氨基酸序列分别为DWGHSW、VFYPW、GTGW和TYFPHF；其中，VFYPW表现出最高的DPPH自由基清除率（IC50：（0.350±0.013）mg/mL）和ABTS阳离子自由基清除率（IC50：（9.482±0.122）µg/mL），其次为DWGHSW；氢键和疏水作用使DWGHSW和VFYPW与Keap1蛋白（PDB ID：2FLU）形成稳定的复合物。该研究可为宁夏肉牛血红蛋白肽作为天然源抗氧化补充剂的应用提供理论依据，有助于宁夏肉牛血液资源的高值化利用。

关键词: 计算机筛选；分子模拟；宁夏肉牛血红蛋白；抗氧化肽；鉴定

Abstract: 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.

Key words: computerized screening; molecular simulation; hemoglobin of Ningxia beef cattle; antioxidant peptides; identification

中图分类号:

TS251.93

徐斐然，王玉，王晓静，崔伟，王永坤，周辉，王兆明，蔡克周，章宝，徐宝才. 基于计算机筛选与分子模拟技术的宁夏肉牛血红蛋白源抗氧化肽的鉴定[J]. 食品科学, 2025, 46(2): 108-117.

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[J]. FOOD SCIENCE, 2025, 46(2): 108-117.

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基于计算机筛选与分子模拟技术的宁夏肉牛血红蛋白源抗氧化肽的鉴定

Hemoglobin-Derived Antioxidant Peptides from Beef Cattle in Ningxia: Identification Based on Computerized Screening and Molecular Docking

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