Synthesis and Use of New Semispecific Substrates for Trypsin-Catalyzed Peptide Bond Formation

Abstract

Two series of semispecific acyl donors, hydroxyalkyl esters of Z-Ala-OH and TV-modified carboxamidomethyl (Cam) esters of Z-Xaa-OH (Xaa = Ala, Leu, Phe) were synthesized as substrates for trypsin-catalyzed peptide synthesis. It follows from the specificity constants of these compounds, that the carboxamidomethyl derivatives are well accepted by trypsin due to favourable S2′ – P2′ interactions. These new substrates can be successfully used for the trypsin-mediated formation of dipeptide amides. The synthesis outcome depends on the amino acid in the P1 position, the ability of the leaving group to provide efficient interactions with the enzyme subsite and the hydrophobicity of the nucleophilic amino acid amide. The modified Cam esters give better peptide yields in comparison to the unmodified ones.