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.
出处
《天然产物研究与开发》
CAS
CSCD
北大核心
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)
关键词
香椿子
亲和层析
蛋白纯化
抗氧化
Toona sinensis seeds affinity chromatography protein purification antioxidant activity
