摘要
我们用正丁醇抽提，经ＳｅｐｈａｄｅｘＧ－１００凝胶过滤和ＤＥＡＥ－纤维素柱层析两步纯化，得到了比活性增大４０倍和ＰＡＧＥ纯的牛ＡＫＰ同工酶。对各ＡＫＰ同工酶分别进行酶活性、电泳迁移率（Ｒｆ值）、热抑制、尿素抑制、组织特异性抑制实验，以鉴别每种同工酶的特性。实验结果表明：①各同工酶Ｒｆ值，肝同工酶＞骨同工酶＞小肠同工酶；②经５６℃加热１０ｍｉｎ，只有骨同工酶完全失活，而肠同工酶对热是稳定的；③尿素强烈地抑制骨同工酶活性；肠同工酶对尿素是稳定的；④组织特异性抑制剂苯丙氨酸强烈抑制肠ＡＫＰ活性。
Alkaline phosphatase isoenzymes with a specific activity of >40 fold and PAGE pure was isolated from the liver,kidney,bone and intestine of the newborn calf (First,each tissue homogentes was extracted in buffer butanol Then,the aqueous phase was separated by filtration on Sephadex G-100,DEAE-cellulose column chromatography) The pure product was studied with respect to its enzymetic activity,electrophoretic mobilities ( Rf value),heat inhibition,urea inhibition,organ-specific inhibitor treatment,to establish the identities of each isoenzyme The results demonstrated the following: 1 Isoenzymes Rf value:liver isoenzyme>bone isoenzyme>small intestine isoenzyme 2 Only the bone enzyme was completely inactivated when heated at 56℃ 10 min Intestial isoenzyme was heat-stable 3 Urea was a strict inhibitor of bone isoenzyme and intestial isoenzyme was urea-stable 4 Organ-specific inhibitor strictly inhibit activity of intestine AKP
关键词
牛
碱性磷酸酶
同工酶
特性
Alkaline phosphatase isoenzyme, Electrophoretic mobilities, Heat inhibition, Urea inhibition, Organ-specific inhibition
