Fujiwara K, Tsuru D
J Biochem. 1978 Apr;83(4):1145-9.
L-Pyroglutamyl derivatives of p-nitroaniline and 7-amino-4-methylcoumarin were synthesized as new sensitive substrates for pyrrolidonyl peptidase (pyrrolidonecarboxylyl peptidase) from Bacillus amyloliquefaciens. Their hydrolyses could be followed by conventional colorimetric and fluorometric procedures; i.e., in terms of the increase in absorbance at 410 nm caused by the liberation of p-nitroaniline and the emission at 440 nm after excitation at 370 nm depending on the liberation of 7-amino-4-methylcoumarin. Values of Km were estimated to be 0.69 mM for anilide substrate and 0.33 mM for methylcoumarin substrate in the pyrrolidonyl peptidase reaction at pH 8.0. The methylcoumarin compound was about one thousand fold more sensitive than the anilide substrate.
合成了对硝基苯胺和7-氨基-4-甲基香豆素的L-焦谷氨酸衍生物，作为解淀粉芽孢杆菌中吡咯烷酮基肽酶（吡咯烷酮羧基肽酶）的新型灵敏底物。它们的水解反应可以通过常规的比色法和荧光法进行监测；也就是说，根据对硝基苯胺的释放导致410nm处吸光度的增加，以及根据7-氨基-4-甲基香豆素的释放，在370nm激发后440nm处的发射。在pH 8.0的吡咯烷酮基肽酶反应中，苯胺底物的Km值估计为0.69 mM，甲基香豆素底物的Km值为0.33 mM。甲基香豆素化合物的灵敏度比苯胺底物高约一千倍。
