Peptide bond formation by aminolysin-A catalysis: A simple approach to enzymatic synthesis of diverse short oligopeptides and biologically active puromycins

Hirokazu Usuki, Yukihiro Yamamoto, Jiro Arima, Masaki Iwabuchi, Shozo Miyoshi, Teruhiko Nitoda, Tadashi Hatanaka

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

A new S9 family aminopeptidase derived from the actinobacterial thermophile Acidothermus cellulolyticus was cloned and engineered into a transaminopeptidase by site-directed mutagenesis of catalytic Ser491 into Cys. The engineered biocatalyst, designated aminolysin-A, can catalyze the formation of peptide bonds to give linear homo-oligopeptides, hetero-dipeptides, and cyclic dipeptides using cost-effective substrates in a one-pot reaction. Aminolysin-A can recognize several C-terminal-modified amino acids, including the l- and d-forms, as acyl donors as well as free amines, including amino acids and puromycin aminonucleoside, as acyl acceptors. The absence of amino acid esters prevents the formation of peptides; therefore, the reaction mechanism involves aminolysis and not a reverse reaction of hydrolysis. The aminolysin system will be a beneficial tool for the preparation of structurally diverse peptide mimetics by a simple approach.

Original languageEnglish
Pages (from-to)2327-2335
Number of pages9
JournalOrganic and Biomolecular Chemistry
Volume9
Issue number7
DOIs
Publication statusPublished - Apr 7 2011

Fingerprint

Puromycin
Oligopeptides
Catalysis
catalysis
peptides
amino acids
Dipeptides
Amino Acids
Peptides
thermophiles
synthesis
Puromycin Aminonucleoside
mutagenesis
Mutagenesis
Aminopeptidases
Site-Directed Mutagenesis
Amines
hydrolysis
esters
Hydrolysis

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Biochemistry

Cite this

Peptide bond formation by aminolysin-A catalysis : A simple approach to enzymatic synthesis of diverse short oligopeptides and biologically active puromycins. / Usuki, Hirokazu; Yamamoto, Yukihiro; Arima, Jiro; Iwabuchi, Masaki; Miyoshi, Shozo; Nitoda, Teruhiko; Hatanaka, Tadashi.

In: Organic and Biomolecular Chemistry, Vol. 9, No. 7, 07.04.2011, p. 2327-2335.

Research output: Contribution to journalArticle

Usuki, Hirokazu ; Yamamoto, Yukihiro ; Arima, Jiro ; Iwabuchi, Masaki ; Miyoshi, Shozo ; Nitoda, Teruhiko ; Hatanaka, Tadashi. / Peptide bond formation by aminolysin-A catalysis : A simple approach to enzymatic synthesis of diverse short oligopeptides and biologically active puromycins. In: Organic and Biomolecular Chemistry. 2011 ; Vol. 9, No. 7. pp. 2327-2335.
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