Evaluation of the specificity of cardosin in peptide synthesis

Marlene Barros, Euclides Fifes

Research output: Contribution to journalArticlepeer-review

Abstract

In the growing field of preparative synthesis of peptides, enzymatic peptide synthesis has become an integrated part of the synthesis valuable as a synthesis technique and as a too) for fragment condensations. Recently in our lab it was described the isolation and characterization of a new enzyme, an aspartate proteinase (cardosin) that exhibits a high specificity for bonds between hydrophobic amino acids and is stable in aqueous-organic biphasic systems. In this work the specificity of cardosin in relation to the amino acids in PI and P'1 position in terms of peptide bond synthesis was investigated and the results were compared with pepsin under the same conditions. In what concern the secondary specificity of cardosin we evaluate the effect of Val in P2 position. The condensation reactions were carried out in aqueousorganic biphasic systems of n-hexane/ethyl acetate and sodium phosphate buffer. The reaction products were isolated by RP-HPLC and identified by amino acids analysis and M.S. Under the conditions described cardosin showed the following preference about P'l position: Met>Phe>Val, but pepsin followed the order: Phe>Met>Val. The cardosin's PI position preference followed the order: CBZ.Phe>CBZ.Leu. The production of tripeptides by condensation of CBZ.Val.Phe with Phe.OMe, Met.OMe and Val.OMe allows us to verify the effect of the amino acid Val on the position P2. The results shows that the addition of Val in the P2 position control the cardosin preference in relation to the amino acid in P l position. Cardosin with the characteristics in terms of specificity, stability in the presence of organic solvents and the facility in their acquisition and purification, can be a new enzyme available for enzymatic peptide synthesis.
Original languageEnglish
Pages (from-to)A974
JournalFASEB Journal
Volume10
Issue number6
Publication statusPublished - 1996
Externally publishedYes

Fingerprint

Dive into the research topics of 'Evaluation of the specificity of cardosin in peptide synthesis'. Together they form a unique fingerprint.

Cite this