A novel L-amino acid ligase from Bacillus subtilis NBRC3134, a microorganism producing peptide-antibiotic rhizocticin

Kuniki Kino, Yoichi Kotanaka, Toshinobu Arai, Makoto Yagasaki

Research output: Contribution to journalArticlepeer-review

42 Citations (Scopus)

Abstract

L-Amino acid ligase catalyzes the formation of an α-peptide bond from unprotected L-amino acids in an ATP-dependent manner, and this enzyme is very useful in efficient peptide production. We performed enzyme purification to obtain a novel L-amino acid ligase from Bacillus subtilis NBRC3134, a microorganism producing peptide-antibiotic rhizocticin. Rhizocticins are dipep-tide or tripeptide antibiotics and commonly possess L-arginyl-L-2-amino-5- phosphono-3-cis-pentenoic acid. The purification was carried out by detecting L-arginine hydroxamate synthesis activity, and a target enzyme was finally purified 1,280-fold with 0.8% yield. The corresponding gene was then cloned and designated rizA. rizA was 1,242 bp and coded for 413 amino acid residues. Recombinant RizA was prepared, and it was found that the recombinant RizA synthesized dipeptides whose N-terminus was L-arginine in an ATP-dependent manner. RizA had strict substrate specificity toward l-arginine as the N-terminal substrate; on the other hand, the substrate specificity at the C-terminus was relaxed.

Original languageEnglish
Pages (from-to)901-907
Number of pages7
JournalBioscience, Biotechnology and Biochemistry
Volume73
Issue number4
DOIs
Publication statusPublished - 2009

Keywords

  • L-amino acid ligase
  • Peptide synthesis
  • Rhizocticin

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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