Hergenrother P J, Haas M K, Martin S F
Department of Chemistry and Biochemistry, University of Texas at Austin 78712, USA.
Lipids. 1997 Jul;32(7):783-8. doi: 10.1007/s11745-997-0101-5.
A rapid and convenient chromogenic assay for phospholipase D from Streptomyces chromofuscus (PLDSc) has been developed that converts the choline generated from the enzyme-catalyzed hydrolysis of phospholipids into a chromogenic dye. By quenching the reaction with EDTA at defined times, an initial rate curve is produced from which a kcat and K(m) can be readily derived. This assay has been applied to the biological evaluation of several substrate analogs, all of which appear to be activators rather than substrates or inhibitors of this enzyme. Performing the assay in 96-well microtiter plates allows for the easy screening of potential effectors of this enzyme.
已开发出一种快速便捷的生色测定法，用于检测来自暗褐链霉菌的磷脂酶D（PLDSc），该方法可将酶催化磷脂水解产生的胆碱转化为生色染料。通过在特定时间用EDTA淬灭反应，可生成初始速率曲线，从中可轻松得出kcat和K(m)。该测定法已应用于几种底物类似物的生物学评估，所有这些类似物似乎都是该酶的激活剂，而非底物或抑制剂。在96孔微量滴定板中进行该测定，便于对该酶的潜在效应物进行筛选。
