人UGT1A3重组酶催化芹菜素葡醛酸结合反应  

Glucuronidation of apigenin by the recombinant human UGT1A3

作　　者：谢升谷[1] 陈亚坤[1] 陈枢青[1] 曾苏[1] 

机构地区：[1]浙江大学药学院,浙江杭州310031

出　　处：《中国药理学与毒理学杂志》2006年第5期405-409,共5页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金资助项目(30100232);国家自然科学基金资助项目(30225047)~~

摘　　要：目的旨在了解人UGT1A3重组酶与芹菜素的有关代谢及其酶动力学参数,并阐述不同有机溶剂对酶动力学参数测定的影响。方法采用Bac-to-Bac系统表达人UGT1A3重组酶与芹菜素37℃共孵育,用HPLC法测定孵育液中剩余底物浓度,利用Lineweaver Burk法计算酶动力学参数,并进一步进行代谢物的确认。同时考察了用不同有机溶剂溶解底物对酶动力学参数测定的影响。结果采用Bac-to-Bac系统表达人UGT1A3重组酶,测得其催化芹菜素的Km为(28.88±2.47)μmol.L-1,Vmax为(224.19±21.11)nmol.m in-1.g-1,Vmax/Km为(7.75±0.29)mL.m in-1.g-1。不同有机溶剂对酶动力学参数测定无显著影响。结论芹菜素可被人UGT1A3重组酶催化,进行葡醛酸结合反应。AIM In order to explore the relationship between human UGT1A3 and apigenin metabolism, its kinetic parameters, and the effects of organic solvents. METHODS The Bacto-Bac expression system was used to express the recombinant UGT1A3. The enzyme activity towards apigenin in cell homogenate protein was measured by HPLC. The kinetic parameters effected by organic solvents were also measured by the same methods. The metabolite was validated by β-glucuronidase and mass chromatogram. RESULTS UGT1A3 activity towards apigenin in cell homogenate protein was found to be Km （28.88 ± 2.47 ） μmol·L^ -1 , Vmax （224. 19 ±21. 11）nmol·min^-1 ·g^-1, Vmax/Km（7.75±0.29）mL·min^-l·g^-1（n=3）. The organic solvents generally had a relatively minor effect on the enzyme activity. CONCLUSION UGT1A3 catalyzed the glucuronidation of apigenin, and organic solvents have a minor affect on UGT1A3 activity.

关 键 词：UGT1A3重组酶 芹菜素 葡醛酸结合反应 动力学参数 有机溶剂影响 

分 类 号：R914[医药卫生—药物化学]