Papain-Specific Activating Esters in Aqueous Dipeptide Synthesis

• Published in: Chembiochem : a European journal of chemical biology 2012-06, Vol.13 (9), p.1319-1326
• Main Authors: de Beer, Roseri J. A. C., Zarzycka, Barbara, Mariman, Michiel, Amatdjais-Groenen, Helene I. V., Mulders, Marc J., Quaedflieg, Peter J. L. M., van Delft, Floris L., Nabuurs, Sander B., Rutjes, Floris P. J. T.
• Format: Article
• Language: English
• Subjects: Biocatalysis; Catalytic Domain; Chemistry Techniques, Synthetic; Dipeptides - chemical synthesis; Dipeptides - chemistry; Drug Design; enzymatic peptide synthesis; enzyme-specific activation; Esters; Models, Molecular; molecular modeling; papain; Papain - chemistry; Papain - metabolism; peptides; Water - chemistry
• ISSN: 1439-4227; 1439-7633
• DOI: 10.1002/cbic.201200017

Enzymatic peptide synthesis has the potential to be a viable alternative for chemical peptide synthesis. Because of the increasing commercial interest in peptides, new and improved enzymatic synthesis methods are desirable. In recently developed enzymatic strategies such as substrate mimetic approaches and enzyme‐specific activation, use of the guanidinophenyl ester (OGp) group has been shown to suffer from some drawbacks. OGp esters are sensitive to spontaneous chemical hydrolysis and the group is expensive to synthesize and therefore not suitable for large‐scale applications. On the basis of earlier computational studies, we hypothesized that OGp might be replaceable by simpler ester groups to make the enzyme‐specific activation approach to peptide bond formation more accessible. To this end, a set of potential activating esters (Z‐Gly‐Act) was designed, synthesized, and evaluated. Both the benzyl (OBn) and the dimethylaminophenyl (ODmap) esters gave promising results. For these esters, the scope of a model dipeptide synthesis reaction under aqueous conditions was investigated by varying the amino acid donor. The results were compared with those obtained from a previous study of Z‐XAA‐OGp esters. Computational docking analysis of the set of esters was performed in order to provide insight into the differences in the reactivities of all the potential activating esters. Finally, selected ODmap‐ and OBn‐activated amino acids were applied in the synthesis of two biologically active dipeptides on preparative scales. A set of new activating esters was evaluated in an enzymatic, papain‐catalyzed approach to synthesizing dipeptides under aqueous conditions. In particular, benzyl and dimethylaminophenyl esters yielded the corresponding dipeptides in several instances in high yields and with relatively small levels of hydrolysis. These results were also interpreted by computational docking analysis.