Department of Biomedical Sciences and Informatics, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Kyoto 610-0394, Japan.
Molecules. 2021 Jan 18;26(2):482. doi: 10.3390/molecules26020482.
A molecular probe with l-phenylalanine -nitroanilide and l-lysin 4-methylcoumaryl-7-amide, in which these amino acid derivatives are connected through a succinic-acid spacer, was prepared. Trypsin and papain were detected by blue-fluorescence emission of generated 7-amino-4-methylcoumarin (AMC). α-Chymotrypsin and nattokinase were detected from both the blue-fluorescence emission of AMC and the UV absorbance of -nitroaniline. In addition, different time courses of -nitroaniline and AMC were observed between the reaction of with α-chymotrypsin and that with nattokinase. In the case of nattokinase, both the fluorescence emission and UV absorbance slowly increased. In contrast, the increasing UV absorbance was saturated at the early stage of the reaction of the present probe with chymotrypsin, whereas the fluorescence emission continuously increased in the following stages.
制备了一种分子探针，其结构为 l-苯丙氨酸-硝基苯胺和 l-赖氨酸 4-甲基香豆素-7-酰胺，其中这些氨基酸衍生物通过琥珀酸间隔基连接。通过生成的 7-氨基-4-甲基香豆素（AMC）的蓝色荧光发射来检测胰蛋白酶和木瓜蛋白酶。通过 AMC 的蓝色荧光发射和 -硝基苯胺的紫外吸收来检测α-糜蛋白酶和纳豆激酶。此外，观察到与α-糜蛋白酶的反应与与纳豆激酶的反应之间 -硝基苯胺和 AMC 的不同时间进程。在纳豆激酶的情况下，荧光发射和紫外吸收均缓慢增加。相比之下，在与糜蛋白酶的反应的早期阶段，本探针的 UV 吸收增加达到饱和，而荧光发射在随后的阶段继续增加。
