松仁谷蛋白抗氧化肽的分离纯化及一级结构鉴定  

Isolation,Purification and Structural Analysis of Antioxidant Peptides Derived from Gluten in Pinus koraiensis Sieb.et Zucc Nut Kernels

作　　者：毛晶[1] 王鹏[1] 王祖浩[1] 方丽[1] 王辑[1] 闵伟红[1] MAO Jing;WANG Peng;WANG Zuhao;FANG Li;WANG Ji;MIN Weihong(National Engineering Laboratory on Wheat and Corn Further Processing, College of Food Science and Engineering,Jilin Agricultural University, Changchun 130118, China)

机构地区：[1]吉林农业大学食品科学与工程学院,小麦和玉米深加工国家工程实验室,吉林长春130118

出　　处：《食品科学》2017年第3期59-63,共5页Food Science

基　　金：国家高技术研究发展计划(863计划)项目(2013AA102206)

摘　　要：利用碱性蛋白酶酶解制备松仁谷蛋白水解物,采用超滤、Sephadex G-25、Sephadex G-15及反相高效液相色谱对酶解物进行分离纯化,并利用质谱进行结构鉴定。对质谱结果进行从头测序,筛选得到两种抗氧化肽肽段Glu-Leu-Leu-Lys-Leu-His(ELLKLH)、Glu-Lys-Asp-Phe-His-Leu(EKDFHL)。经固相合成后得到纯度分别为99.67%和95.15%的肽段,对其进行抗氧化活性测定。结果表明,在浓度为1 000 μmol/m L时,ELLKLH和EKDFHL的1,1-二苯基-2-三硝基苯肼自由基清除率分别为38.26%和29.54%,铁离子螯合率分别为56.44%和48.66%。In this work, the enzymatic hydrolysate of pine nut gluten produced with alkaline protease was isolated and purified by ultrafiltration, Sephadex G-25, Sephadex G-15 and reversed-phase high-performance liquid chromatography (RP-HPLC). The structural characteristics of the purified fractions were identified by Q Exactive hybrid quadrupole-Orbitrap mass spectrometry, and de novo sequencing was performed based on the mass spectral data. Two antioxidant peptides were obtained with amino acid sequence of Glu-Leu-Leu-Lys-Leu-His (ELLKLH) and Glu-Lys-Asp-Phe-His-Leu (EKDFHL),respectively. After solid-phase synthesis, the purity of the peptides was 99.67% and 95.15%, respectively. The results of antioxidant activities showed that the DPPH free radical scavenging rates of ELLKLH and EKDFHL at a concentration of 1 000 μmol/mL were 38.26% and 29.54%, respectively, and the iron ion chelating rates were 56.44% and 48.66%, respectively.

关 键 词：松仁谷蛋白 分离纯化 结构鉴定 

分 类 号：TS209[轻工技术与工程—食品科学]