血红素亲和层析快速纯化香椿子抗氧化活性蛋白  

Rapid Purification of Antioxidant Proteins from Toona sinensis Seeds Using Affinity Chromatography

作　　者：王荣申[1] 孟超[1] 张守军[1] 卢燕[1] 李万忠[1] 

机构地区：[1]潍坊医学院药学院,潍坊261053

出　　处：《天然产物研究与开发》2017年第1期96-100,86,共6页Natural Product Research and Development

基　　金：国家自然科学基金(81274049)

摘　　要：以蛋白质含量、总还原力和DPPH清除能力为指标,对盐析法、碱提酸沉法、直接加热法进行评价,选取蛋白含量高同时有较强还原力的碱提酸沉提取法。将环氧氯丙烷活化血红素偶联于Sephadex G-25制作亲和层析柱,以水为平衡液、Na Ac-HAc缓冲液为洗脱液进行香椿子蛋白纯化,考马斯亮蓝法测定所得蛋白纯度,并对其进行初步抗氧化研究。结果表明:目标产物蛋白具有较好的DPPH清除能力,含量由59.51%提高至96.23%,纯化后蛋白纯度得到显著提高(P<0.05)。Toona sinensis seeds protein （TSP）was extracted by salting out,alkali extraction and acid precipitation,direct heating methods.The total antioxidant and DPPH free radicals scavenge activity of the extract were then evaluated.TSP from alkali extraction and acid precipitation was selected as purification sampleand was purificated by affinity chromatography.Sephadex G-25 were activated using epichlorohydrin,hemin was bound with them to prepare an immobilized hemin affinity chromatography column,which was used to purifyproteins from T.sinensis seeds equilibrated with water and eluted with NaAc-HAc buffer.Purification protein was tested by DPPH free radicals scavenge assay.The results showed that protein content was from 59.51% to 96.23% after affinity chromatography purification.

关 键 词：香椿子 亲和层析 蛋白纯化 抗氧化 

分 类 号：R932[医药卫生—生药学]