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Purification and characterization of an antioxidant protein (~16 kDa) from Terminalia chebula fruit

机译:Terminal子果实中抗氧化蛋白(〜16 kDa)的纯化与鉴定

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Terminalia chebula fruit is used as folk medicine in India and Southeast Asia. An antioxidant protein was isolated by bioassay guided fractionation of I. chebula fruit by homogenizing in the citrate phosphate buffer. The isolated protein (TCP-Ill) obtained from fruit was purified by gel chromatography and preparative HPLC, showed apparent molecular weight of 16 kDa by SDS-PAGE and MALDI-TOF/MS analyses. Amino acid sequence obtained by LC-MS~E analysis showed homology with the predicted protein fragments of Populus trichocarpa, putative uncharacterized protein fragments from Oryza sativa and with fragments of 17 kDa thylakoid lumenal protein from Spinacia oleracea. TCP-III exhibited significant radical scavenging in DPPH, NO, H_2O_2 and ABTS assays. In addition, TCP-III inhibited oxidation of linoleic acid in p-car-otene bleaching assay, reduced ferric ions and chelated ferrous ions. The present finding demonstrates uniquely, for the first time, characterization of an antioxidant protein from T. chebula fruit.
机译:Terminalia chebula果实在印度和东南亚用作民间药。通过在柠檬酸盐磷酸盐缓冲液中匀浆,通过生物测定指导的I. Chebula水果分级分离抗氧化剂蛋白。通过凝胶色谱和制备型HPLC纯化获自水果的分离的蛋白质(TCP-III),通过SDS-PAGE和MALDI-TOF / MS分析显示表观分子量为16kDa。通过LC-MS_E分析获得的氨基酸序列与毛果杨的预测蛋白片段,水稻的推定的未表征的蛋白片段和菠菜的17kDa类囊体腔蛋白片段具有同源性。在DPPH,NO,H_2O_2和ABTS分析中,TCP-III表现出明显的自由基清除作用。另外,TCP-III在对胡萝卜素漂白试验中抑制了亚油酸的氧化,减少了铁离子和螯合的亚铁离子。本发现首次独特地证明了来自T. chebula果实的抗氧化剂蛋白的表征。

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