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The applicability of subtilisin carlsberg in peptide synthesis
Author(s) -
Uwe Klein Jens,
Prykhodzka Alexandra,
Čeřovský Václav
Publication year - 2000
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/1099-1387(200011)6:11<541::aid-psc271>3.0.co;2-t
Subject(s) - chemistry , nucleophile , tripeptide , subtilisin , peptide , acylation , amino acid , peptide synthesis , acetonitrile , stereochemistry , peptide bond , dipeptide , glycine , pyridine , hydrolysis , organic chemistry , medicinal chemistry , enzyme , biochemistry , catalysis
The synthesis of peptide bonds catalysed by subtilisin Carlsberg was studied in different hydrophilic organic solvents with variable H 2 O concentration. Z‐Val‐Trp‐OMe and Z‐Ala‐Phe‐OMe were used as acyl donors, and a series of amino acid derivatives, di‐ and tripeptides of the general structure Xaa‐Gly, Gly‐Xaa, Gly‐Gly‐Xaa (Xaa represents all natural L‐amino acids except cysteine) and other peptides were used as nucleophiles. A comparative study of the enzymatic synthesis in aqueous DMF (50%, v/v) and acetonitrile containing 10% (v/v) of H 2 O demonstrated that the yields of peptide products were higher in most cases when acetonitrile with low H 2 O concentration was used. The acylation of weak nucleophiles was improved in organic solvents with very low H 2 O concentration (2%). The reactions in anhydrous Bu t ‐OH proceeded with substantially lower velocity. Generally, the restricted nucleophile specificity of the enzyme for glycine and hydrophilic amino acid residues in P1′ position, as well as numerous side reactions, limit the utilization of subtilisin in peptide synthesis, especially in the case of the segment condensations. Contrary to the published data, we have proved that proline derivatives were not acylated in any media with the help of subtilisin Carlsberg. Effective ester hydrolysis of a protected nonapeptide corresponding to the N‐terminal sequence of dicarba‐eel‐calcitonin catalysed by sublilisin was achieved. Copyright © 2000 European Peptide Society and John Wiley & Sons, Ltd.