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《Chinese Archives of Traditional Chinese Medicine》 2011-05
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Study of the Isolation and Purification of Antioxidative Peptides from Short Necked Clam,Ruditapes Philippinarum

YANG Yong-fang,YANG Zui-su,DING Guo-fang,YU Di,HUANG Fang-fang(School of Food Science and Pharmacy of Zhejiang Ocean University,Zhoushan 316004,Zhejiang,China)  
Objective:In order to get products with antioxidant activity from Ruditapes philippinarum.Methods:Four proteases(pepsin,trypsin,papain,alcalase) were applied to hydrolyze the R.philippinarum.The antioxidative activity of the fractions against the hydroxyl radical produced by H2O2/Fe2+ was determined in vitro.Ultrafiltration and anion-exchange chromatography and reversed-phase high-performance liquid chromatography(RP-HPLC) were performed to separate and purify the antioxidant peptides.Results:Hydrolysate with trypsin had the highest antioxidant activity and was fractionated by ultrafiltration.Three fractions(3KDa,3~5KDa,and5KDa) were separated.Below 3KDa fraction,which exhibited the highest antioxidative activity,was further purified using anion-exchange and reverse phase high performance liquid chromatography.An antioxidative peptide was isolated and its clearance rate of hydroxyl radical and reducing power were higher than vitamin C.The purified peptide sequences with a molecular weight of 607.6Da was identified as Asp-Trp-Pro-His.Conclusion:Products hydrolyzed by trypsin from R.philippinarum exhibited the best antioxidant activity among all proteases selected,and could be isolated and purified by the methods of ultrafiltration and anion-exchange chromatography and RP-HPLC.
【Fund】: 浙江省自然科学基金资助项目(Y3100129);; 浙江省教育厅和浙江省财政厅研究生创新科研项目(YK2008072);浙江省教育厅资助项目(Y200907990);; 舟山市科技计划项目(092016)
【CateGory Index】: R285.5
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