Iusupova M P, Kotlova E K, Timokhina E A, Stepanov V M
Bioorg Khim. 1995 Jan;21(1):33-8.
Subtilisin 72 sorbed on the surface of macroporous glass catalyzes a condensation of the esters of N-acylated peptides with arginine derivatives in organic solvents. The sorbed enzyme can be used repeatedly, which makes it possible to synthesize the chromophore substrates of metalloproteinases and carbopeptidases of the general formula Dnp-Ala-Ala-Xaa-Arg-NH2 (Xaa = Leu, Phe, Val, Ile). In tetrapeptides, metalloproteinases hydrolyze the Ala-Xaa bond with the removal of Dnp-Ala-Ala-OH, which can be determined spectrophotometrically. The chromophore substrates of carboxypeptidases of the B type (Dnp-Ala-Ala-Xaa-Arg-OH and Dnp-Ala-Ala-Arg-OH) are obtained by hydrolysis of the corresponding amides by trypsin.
吸附在大孔玻璃表面的枯草杆菌蛋白酶72可催化N-酰化肽酯与精氨酸衍生物在有机溶剂中的缩合反应。吸附的酶可重复使用，这使得合成通式为Dnp-Ala-Ala-Xaa-Arg-NH2（Xaa = Leu、Phe、Val、Ile）的金属蛋白酶和羧肽酶的发色底物成为可能。在四肽中，金属蛋白酶水解Ala-Xaa键，同时去除Dnp-Ala-Ala-OH，后者可通过分光光度法测定。B型羧肽酶的发色底物（Dnp-Ala-Ala-Xaa-Arg-OH和Dnp-Ala-Ala-Arg-OH）是通过胰蛋白酶水解相应的酰胺得到的。
