【摘要】 用硫酸铵分级沉淀、ＤＥ５２、ＳｅｐｈａｄｅｘＧ－１５０、ＤＥＡＥ－ＳｅｐｈａｄｅｘＡ－５０及Ｐｈｅｎｙｌ－ＳｅｐｈａｒｏｓｅＣＬ－４Ｂ柱层析，纯化获得电泳纯人肝ＧＳＨｐｘ。经最后一步ＤＥＡＥ－ＳｅｐｈａｄｅｘＡ－５０柱层析，酶活性峰和蛋白峰吻合，比活性为６５５ｕ／ｍｇ蛋白，纯化４２６２倍，产率９．８％。该酶的ＳＤＳ－ＰＡＧＥ显示分子量为２３０００ｋＤ单一区带。免疫家免得抗血清效价为１：６４，琼脂双向扩散试验见人肝和红细胞ＧＳＨ－ｐｘ间发生交叉免疫反应。人肝ＧＳＨｐｘ活性最适ｐＨ：在８．５左右；在ｐＨ７．４、８．０及９．０，３７℃环境下２０分钟其活性丢失近８０％，在ｐＨ８．５，３７℃，１００分钟仅丢失３０％。更多还原

【Abstract】 lutathione peroxidase(GSHpx)was purified frotn human liver by(NH4 )2SO4 precipitaion,ED52,sephadex G-150 ,DEAE-sephadex A-50 and phenyl-sepharose CL-4B column chromatography. Through finalDEAE-sephadex A-50 column , the enzyme activity peak and protein peak were entirely overloped in elutionprofile.The enzyme was purified 4 262-fold with a yield of 9.8%and specific activity of 665 U/mg protein.SDS-PAGE of the enzyme showed a single band with molecular weight of 23000 kD.Rabbit antiserum raisedagainst human liver GSHpx was prepared and the antisetum titer measured by Ouchterlony double diffusiontest was 1 :64.The test also revealed a crossimmuno-reaction between human liver and erythrocytes GSHpx.In vitro,the enzyme had a optimal pH at 8.5.About 20%of the original activity was retained after incubationat 37 ℃ for 20 min at pH 7.4,8.0 or 9.0,but 70%for 100 min at pH 8.5.更多还原