血红素肽-铁的分离纯化及其清除自由基能力  

Separation and Purification of Heme Peptide-Iron and Its Capacity of Scavenging Free Radical

作　　者：张滨[1,2] 马美湖[3] 

机构地区：[1]湖南农业大学食品科技学院,长沙410128 [2]长沙环境保护职业技术学院,长沙410004 [3]华中农业大学食品科技学院,武汉430070

出　　处：《中国生物制品学杂志》2010年第1期30-35,共6页Chinese Journal of Biologicals

基　　金：国家"十一五"重大科技专项课题(2006BAD05A16);湖南省教育厅课题(08D012)

摘　　要：目的分离纯化血红素肽-铁,并分析其清除自由基的能力。方法以聚丙烯酰胺为载体,对利用真空及低压高频脉冲胰蛋白酶降解猪血红蛋白的产物进行金属螯合亲和层析分离纯化,利用基质辅助激光解析串联飞行时间质谱(MALDI-TOFMS)及生物信息学网站分析血红素肽-铁的分子结构;以维生素C(Vc)为对照,分析血红素肽-铁的抗氧化能力;以邻苯三酚法分析血红素肽-铁清除超氧阴离子的能力;以脱氧核糖法分析血红素肽-铁清除羟基自由基(·OH)的能力;以三氯甲烷-甲醇法分析血红素肽-铁对脂质过氧化的抑制作用。结果分离纯化的血红素肽-铁由139个氨基酸组成,相对分子质量为16065.08,铁离子含量为11013.5μg/L,理论pI为8.10,消光系数为6990,脂肪系数为92.73,不稳定系数为1.18,总平均疏水性为-0.023;在体外具有较强的清除超氧阴离子和抗氧化自由基能力。结论分离纯化的血红素肽-铁具有清除自由基的能力。Objective To separate and purify heme peptide-iron and analyze its capacity of scavenging free radical.Methods The lysate of porcine haematoglobin with trypsin under conditions of vacuum and low-voltage high frequency pulse was separated and purified by metal chelate affinity chromatography using polyacrylamide as a carrier.The obtained heme peptide-iron was analyzed for molecular structure by MALDI-TOFMS and bioinformatics software,for antioxidation capacity using vitamin C（Vc）as control,for capacity of scavenging superoxide anion by pyrogallol method,for capacity of scavenging hydroxyl radical by deoxyribose （DR）method,and for inhibitory effect on lipid peroidation by chloroform-methanol method.Results The purified heme peptide-iron consisted of 139 amino acids,with a relative molecular mass of 16 065.08,a ferri ion content of 11 013.5 μg /L,a theoretical pI value of 8.10,an extinction coefficient of 6 990,a fat coefficient of 92.73,an instability coefficient of 1.18 and a total mean hydrophobicity of 0.023,and showed high capacity of scavenging superoxide anion and antioxidant in vitro.Conclusion The purified heme peptideiron showed ability in clearance of free radical.

关 键 词：血红素肽-铁 金属螯合亲和层析 生物信息学 自由基 

分 类 号：Q599[生物学—生物化学]