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.