Cloning and characterization of penicillin V acylase from Streptomyces mobaraensis

Demin Zhang, Mayuko Koreishi, Hiroyuki Imanaka, Koreyoshi Imamura, Kazuhiro Nakanishi

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)

Abstract

We report on the molecular cloning and characterization of penicillin V acylase (PVA) from an actinomycete, Streptomyces mobaraensis (Sm-PVA), which was originally isolated as an acylase that efficiently hydrolyzes the amide bond of various N-fatty-acyl-l-amino acids and N-fatty-acyl-peptides as well as capsaicin (8-methyl-N-vanillyl-6-nonenamide). In addition, the purified Sm-PVA hydrolyzed penicillin V with the highest activity (kcat) among the PVAs so far reported, penicillin G, and 2-nitro-5-phenoxyacetamide benzoic acid. The BLAST search revealed that the Sm-PVA precursor is composed of a polypeptide that is characteristic of enzymes belonging to the β-lactam acylase family with four distinct segments; a signal sequence (43 amino acids), an α subunit (173 amino acids), a linker peptide (28 amino acids), and a β subunit (570 amino acids). The mature, active Sm-PVA is a heterodimeric protein with α and β subunits, in contrast to PVAs isolated from Bacillus sphaericus and B. subtilis, which have a homotetrameric structure. The amino acid sequence of Sm-PVA showed identities to PVA from S. lavendulae, N-acylhomoserine lactone-degrading acylase from Streptomyces sp., cyclic lipopeptide acylase from Streptomyces sp., and aculeacin A acylase from Actinoplanes utahensis with 68, 67, 67, and 41% identities, respectively.

Original languageEnglish
Pages (from-to)788-800
Number of pages13
JournalJournal of biotechnology
Volume128
Issue number4
DOIs
Publication statusPublished - Mar 10 2007

Keywords

  • Actinomycete
  • Capsaicin
  • Penicillin V
  • Penicillin V acylase
  • Streptomyces mobaraensis
  • β-Lactam acylase family

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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