Abstract
The kinetically controlled synthesis of N-benzyloxycarbonyl (Z)-dipeptides was investigated by the use of free amino acids as nucleophiles and a cysteine protease papain as catalyst. The coupling efficiency was significantly improved by the combined use of the carbamoylmethyl (Cam) ester of a Z-amino acid as acyl donor and frozen aqueous solution (ice, -16 or -24 °C) as reaction medium. The yield of peptide synthesis became high when both P1- and P′1-positions were occupied by small non-polar amino acids (Z-Gly-Gly-OH, 76%; Z-Gly-Ala-OH, 75%; Z-Ala-Ala-OH, 72%). Similar results were observed by the use of ficin as catalyst instead of papain. Furthermore, this strategy was applied to the papain-catalyzed incorporation of a d-configured amino acid such as d-alanine into the resulting peptides. Although the coupling in aqueous solution (30 °C) afforded the desired Z-dipeptides in low yields, the freezing of reaction medium reduced significantly unfavorable hydrolysis of the acyl donors, resulting in improvement of the coupling efficiency (Z-Gly-d-Ala-OH, 80%; Z-Ala-d-Ala-OH, 45%; Z-d-Ala-Ala-OH, 22%).
Original language | English |
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Pages (from-to) | 537-543 |
Number of pages | 7 |
Journal | Enzyme and Microbial Technology |
Volume | 43 |
Issue number | 7 |
DOIs | |
Publication status | Published - 2008 Dec 10 |
Keywords
- Carbamoylmethyl (Cam) ester
- Ficin
- Free amino acid
- Frozen aqueous solution (ice)
- Papain
- Peptide synthesis
- d-Amino acid
ASJC Scopus subject areas
- Biotechnology
- Bioengineering
- Biochemistry
- Applied Microbiology and Biotechnology