澳洲坚果抗氧化肽的分离纯化及肽段鉴定
Separation,Purification and Identification of Antioxidant Peptides Derived from Macadamia integrifolia
付镓榕 1马尚玄 1魏元苗 1徐文婷 1郭刚军 1贺熙勇1
作者信息
- 1. 云南省热带作物科学研究所,云南景洪 666100;云南省澳洲坚果农业工程研究中心,云南景洪 666100
- 折叠
摘要
为研究澳洲坚果抗氧化肽的抗氧化活性和氨基酸组成,以复配蛋白酶水解澳洲坚果粕制备粗多肽,利用超滤、大孔树脂纯化技术制备了抗氧化活性最佳的分子量小于 1000 Da的多肽,采用Sephadex G-15凝胶对其分离并评价各组分对DPPH、羟基、ABTS+自由基的清除能力与还原能力,筛选出抗氧化活性最强组分,利用液相色谱-串联质谱技术(liquid chromatography and tandem mass spectrometry,LC-MS/MS)进行鉴定并分析.结果表明,葡聚糖凝胶柱层析分离出G1、G2、G3组分,其中G3具有最佳的抗氧化活性,其羟基自由基清除能力半抑制浓度(half maximal inhibitory concentration,IC50)6.18 mg/mL与还原能力IC50 2.19 mg/mL优于谷胱甘肽,DPPH自由基清除能力IC50 0.50 mg/mL,ABTS+自由基清除能力IC50 0.02 mg/mL;通过液相色谱-串联质谱鉴定G3含有 46个肽段,肽段长度均小于 10个氨基酸,匹配得分大于 200分的 9条肽段分子量为 631～920 Da,均无毒性;筛选获得高抗氧化活性肽HLLPK、KEFFP、KEFFPA,其分子量分别为 606.84、666.83、737.92 Da.本研究初步揭示了澳洲坚果抗氧化肽组成,深化了澳洲坚果抗氧化肽的研究,为澳洲坚果抗氧化肽的开发利用提供了理论参考.
Abstract
To research the antioxidant activity and amino acid composition of antioxidant peptides from Macadamia integrifolia,the Macadamia integrifolia polypeptides were prepared by complex protease enzymatic hydrolysis,the cons-tituents with optimal antioxidant activity(molecular weight less than 1000 Da)were obtained by macroporous resin and dialysis technology.The polypeptides were separated by Sephadex G-15 gel,and the scavenging and reducing ability of 1,1-diphenyl-2-picrylhydrazyl(DPPH),hydroxyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate)radicals(ABTS+)of each component were evaluated.The optimal antioxidant components were separated and identified and analyzed by liquid chromatography and tandem mass spectrometry(LC-MS/MS).The results showed that,the G1,G2 and G3 were isolated by dextran gel column chromatography,G3 had the strongest antioxidant activity,its scavenging and reducing capacity of hydroxyl radicals was superior to glutathione with half maximal inhibitory concentration(IC50)of 6.18 mg/mL and 2.19 mg/mL,respectively.The half maximal inhibitory concentration(IC50)of scavenging capacity of DPPH and ABTS+ was 0.50 and 0.02 mg/mL,respectively.G3 was identified by liquid chromatography tandem mass spectrometry to contain 46 peptides,the length of each peptide was less than 10 amino acids.The molecular weights of the 9 peptides with matching scores more than 200 were 631～920 Da,and with no toxicity.The screened peptides with high antioxidant activity were HLLPK,KEFFP and KEFFPA,the molecular weights of them were 606.84,666.83 and 737.92 Da,respectively.This research preliminarily revealed the antioxidant peptides composition of macadamia nut,deepened the research of antioxidant peptide of Macadamia integrifolia,and would provide a theoretical reference for the development and utilization of antioxidant peptide of Macadamia integrifolia.
关键词
澳洲坚果/抗氧化肽/抗氧化活性/分离纯化/肽段鉴定Key words
Macadamia integrifolia/antioxidant peptides/antioxidant activity/separation and purification/peptide identification引用本文复制引用
基金项目
云南省重大科技专项计划(202202AE090006)
云南省盈江县澳洲坚果产业科技特派团(202104BI090004)
云南省热带作物科技创新体系建设专项(RF2024-6)
云南省热带作物科技创新体系建设专项(RF2023-15)
云南省科技人才和平台计划项目(202405AD350060)
兴滇英才支持计划()
西双版纳州"雨林英才支持计划"产业创新人才项目()
出版年
2024
国家科技期刊平台
NSTL
万方数据