CAJ | 학술논문

研究了松仁蛋白分离、纯化、一级结构及部分蛋白清除·OH和O2-·的活性,探讨松仁蛋白的功能机理。松仁蛋白经过硫酸铵盐析初步分级,对收集量最大的分离蛋白P80通过Sephadex G-75凝胶过滤柱和DEAE 52离子交换层析,收集得到P80分离纯化的组分D,对组分D进行基质激光解析质谱分析,结果显示组分D与北美白松松仁清蛋白Albumin 4具有较高的匹配分数,匹配分数为99,分子质量为18647 u,等电点为5.35,序列覆盖率为41%,松仁蛋白组分D可能为与Albumin 4蛋白同类的一种松仁清蛋白或其他蛋白,称其为HSR-Protein 1。体外抗氧化实验表明松仁蛋白具有清除·OH和O2-·的能力。在质量浓度为800 mg/L时,松仁蛋白组分D、松仁蛋白组分P80和松仁粗蛋白对·OH的清除率分别为93.12%、90.89%和89.12%,对O2-·的清除率分别为64.19%、56.45%和35.19%,松仁蛋白组分D对·OH和O2-·的清除能力最强。相同浓度下,纯度越高的松仁蛋白清除·OH和O2-·能力越强。
연구료송인단백분리、순화、일급결구급부분단백청제·OH화O2-·적활성,탐토송인단백적공능궤리。송인단백경과류산안염석초보분급,대수집량최대적분리단백P80통과Sephadex G-75응효과려주화DEAE 52리자교환층석,수집득도P80분리순화적조분D,대조분D진행기질격광해석질보분석,결과현시조분D여북미백송송인청단백Albumin 4구유교고적필배분수,필배분수위99,분자질량위18647 u,등전점위5.35,서렬복개솔위41%,송인단백조분D가능위여Albumin 4단백동류적일충송인청단백혹기타단백,칭기위HSR-Protein 1。체외항양화실험표명송인단백구유청제·OH화O2-·적능력。재질량농도위800 mg/L시,송인단백조분D、송인단백조분P80화송인조단백대·OH적청제솔분별위93.12%、90.89%화89.12%,대O2-·적청제솔분별위64.19%、56.45%화35.19%,송인단백조분D대·OH화O2-·적청제능력최강。상동농도하,순도월고적송인단백청제·OH화O2-·능력월강。
The isolation, purification, primary structure and anti-oxidation of protein from Pinus koraiensis Sieb. et Zucc. seeds were studied and its function mechanism was discussed. The preliminary classification of the protein was conducted by salting out with ammonium sulfate in the filtrate, then the resulting protein ( P80 ) with the highest collection rate was purified by G-75 gel chromatograph and DEAE-cellulose chromatography,and sample D was separated from P80 . Sample D was analyzed by MADIL-TOF-MS. The result showed that sample D and north American white pine nuts Albumin 4 were matched with high scores, and the matched score was 99. And it had the molecular weight of 18 647 u, isoelectric point of 5. 35, and the coverage of gene sequence 41%. Based on the matched score and research results, Albumin 4 and sample D were similar and both a kind of pine cone proteins, entitled HSR-Protein 1. In vitro antioxidant experiment showed that Pinus koraiensis Sieb. et Zucc. seeds protein had good scavenging activities of ·OH and O2-·. At the concentration of 800 mg/L, the ·OH cleaning rate of pine nut protein component D, protein components P80 , and crude protein were 93. 12%, 90. 89% and 89. 12%, respectively. Pine nut protein components D had the strongest scavenging ability of ·OH. At the same concentration, the O2-·scavenging rates of pine nut protein component D, component P80 , and crude protein were 64. 19%, 56. 45% and 35. 19%, respectively. Pine nut protein components D was the strongest. With the same concentration, the scavenging ability became stronger with the purity increasing.