【摘要】 经硫酸铵分级沉淀、ＤＥＡＥ－纤维素离子交换层析、ＳｅｐｈａｄｅｘＧ－２００凝胶柱层析和羟基磷灰石柱层析从Ｒａｊｉ细胞中纯化ＡＤＡ达电泳纯。酶纯化３３６倍，比活力达３５５１５Ｕ／ｍｇ蛋白，回收率为２２８３％；经ＳｅｐｈａｄｅｘＧ－２００凝胶过滤法和ＳＤＳ－ＰＡＧＥ法测得ＡＤＡ的分子量分别为３７０ＫＤ和４２８ＫＤ；以腺苷或脱氧腺苷为底物测得Ｋｍ值分别为１６６６７μｍｏｌ／Ｌ和１０１９５μｍｏｌ／Ｌ．更多还原

【Abstract】 ADA was purified from Raji cells by the procedures of ammonium sulfate differential precipitation,DEAE-cellulose chromatography,Sephadex G-200 gel filtration and hydeoxyapatite chromatography.Compared with the crude extract,the resulting enzyme was purified by 336 fold with 35515 units/mg protein of specific activity and 22.83% of recovery.The purified ADA showed a single band in SDS-PAGE.Its molecular weight is 37.0 KD and 42.8 KD by Sephadex G-200 gel filtration and SDS-PAGE,respectively.When using adenosine and deoxyadenosine as substrates,Km value is 166.67 μ mol/L and 101.95 μmol/L,respectively.更多还原