肉类研究2025，Vol.39Issue(3)：8-17,10.DOI:10.7506/rlyj1001-8123-20240809-203
猪骨源抗氧化肽分离纯化、结构鉴定及抗氧化作用机制
Isolation,Purification,Structural Identification and Action Mechanism of Antioxidant Peptides Derived from Porcine Bones
摘要
Abstract
The enzymatic preparation of antioxidant peptides from porcine bone meal was optimized by the combined use of single factor experiments and response surface methodology.The antioxidant peptides were separated and purified by successive ultrafiltration and Sephadex G-25 column chromatography,and the peptides with high antioxidant activity were selected and identified using liquid chromatography-tandem mass spectrometry(LC-MS/MS)combined with PeptideRanker.The binding of the peptides to the protein Kelch-like ECH-associated protein 1(Keap1)was analyzed by molecular docking technology.The results showed that the optimal process parameters were as follows:pH 6.8,50℃,6 000 U/g flavourzyme,4 h hydrolysis,and 4 g/100 mL substrate concentration.Among three fractions(F-1,F-2,and F-3)obtained from successive ultrafiltration and Sephadex G-25 column chromatography,F-2 had the highest antioxidant activity,which scavenged(94.57±1.08)%of 2,2'-azino-bis(3-ethylbenzothiazoliline-6-sulfonic acid)(ABTS)cationic radical and(79.08±0.61)%of 1,1-diphenyl-2-picrylhydrazyl(DPPH)radical.LC-MS/MS identified 232 peptides from F-2,and three peptides,GPVGAAGP,APGPVGP and QPGPAGPG,were predicted by PeptideRanker,hydrophobic amino acids accounting for more than 80%of their total amino acids,indicating that they may have high antioxidant activity.The three peptides bound to Keap1 protein by hydrogen bonding and hydrophobic interaction,causing the release of nuclear factor erythroid 2 related factor 2(Nrf2)into the nucleus and the activation of the Keap1-Nrf2/antioxidant response element pathway to exert antioxidant effects.Therefore,GPVGAAGP,APGPVGP and QPGPAGPG can be used as new antioxidants,and the findings from this research provide a theoretical basis for the development and utilization of porcine bone-derived antioxidant peptides.关键词
猪骨/酶解/抗氧化肽/分子对接Key words
porcine bones/enzymatic hydrolysis/antioxidant peptides/molecular docking分类
轻工纺织引用本文复制引用
陈露露,雷阳,郝阳雪,吕扬勇,魏闪,赵风光,胡元森..猪骨源抗氧化肽分离纯化、结构鉴定及抗氧化作用机制[J].肉类研究,2025,39(3):8-17,10.基金项目
河南省自然科学基金青年科学基金项目(242300421556) （242300421556）
河南工业大学高层次人才科研启动基金项目(2024BS003) （2024BS003）
横向技术开发项目(062H2023SW062) （062H2023SW062）
