3-脱氧葡萄糖醛酮还原酶药物筛选体系的建立及应用
Establishment and preliminary application of the 3-deoxyglucosone reductase enzymatic drug screening system
作者：江国荣;张露蓉;姚霏;
Author：
收稿日期： 年卷（期）页码：2011,26(05):-441-443
期刊名称：华西药学杂志
Journal Name：WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字：3-脱氧葡萄糖醛酮还原酶;反应体系;药物筛选
Key words：
基金项目：苏州市科技局科教兴卫青年科技项目(SWKQ0821)
中文摘要
目的从鸡肝中提取3-脱氧葡萄糖醛酮(3-DG)还原酶,并建立3-DG还原酶体外反应体系筛选药物。方法以辅酶NADPH的消耗量为评价指标来表示3-DG还原酶的活性,通过对反应体系底物3-DG、辅酶NADPH、酶量和反应时间的优化,考察温度等影响因素,建立3-DG还原酶体外反应体系并应用于寻找提高3-DG还原酶活性的药物。结果优化后的反应体系为:100μg.mL-1 3-DG溶液、250μg.mL-1 NADPH、酶液各20μL,用50 mmol.L-1的Na2HPO4-NaH2PO4缓冲液(pH6.4)调反应体系至200μL,混合物于37℃反应40 min;以NADPH的消耗量来定义3-DG还原酶的酶活单位,并应用于17种药物的筛选。结论该反应体系可用辅酶NADPH的消耗来反映体系中3-DG还原酶的活性,并能为寻找提高3-DG代谢酶活性的药物提供方法。
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