Protease-catalyzed dipeptide synthesis from N-protected amino acid carbamoylmethyl esters and free amino acids in frozen aqueous solutions

Sayed Mohiuddin Abdus Salam, Ken ichi Kagawa, Teruhiko Matsubara, Katsuhiro Kawashiro

Research output: Contribution to journalArticle

15 Citations (Scopus)

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 languageEnglish
Pages (from-to)537-543
Number of pages7
JournalEnzyme and Microbial Technology
Volume43
Issue number7
DOIs
Publication statusPublished - 2008 Dec 10

Fingerprint

Dipeptides
Papain
Amino acids
Esters
Peptide Hydrolases
Amino Acids
Peptides
Ficain
Nucleophiles
Catalysts
Cysteine Proteases
Ice
Freezing
Alanine
Hydrolysis
N-benzyloxycarbonylglycine

Keywords

  • Carbamoylmethyl (Cam) ester
  • d-Amino acid
  • Ficin
  • Free amino acid
  • Frozen aqueous solution (ice)
  • Papain
  • Peptide synthesis

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Protease-catalyzed dipeptide synthesis from N-protected amino acid carbamoylmethyl esters and free amino acids in frozen aqueous solutions. / Salam, Sayed Mohiuddin Abdus; Kagawa, Ken ichi; Matsubara, Teruhiko; Kawashiro, Katsuhiro.

In: Enzyme and Microbial Technology, Vol. 43, No. 7, 10.12.2008, p. 537-543.

Research output: Contribution to journalArticle

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