Recombinant Expression, Biochemical Characterization and Stabilization through Proteolysis of an L-Glutamate Oxidase from Streptomyces sp. X-119-6

Jiro Arima, Takashi Tamura, Hitoshi Kusakabe, Makoto Ashiuchi, Toshiharu Yagi, Hidehiko Tanaka, Kenji Inagaki

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

L-Glutamate oxidase (LGOX) from Streptomyces sp. X-119-6 is a protein of 150 kDa that has hexamer Structure α2β2γ 2. The gene encoding LGOX was cloned and heterologously expressed in Escherichia coli. LGOX isolated from the E. coli transformant had the structure of a one chain polypeptide. Although the recombinant LGOX exhibited catalytic activity, it was inferior to the LGOX isolated from Streptomyces sp. X-119-6 in catalytic efficiency. The recombinant LGOX exhibited low thermostability compared to the LGOX isolated from Streptomyces sp. X-119-6 and was an aggregated form. Proteolysis of the recombinant LGOX with the metalloendopeptidase from Streptomyces griseus (Sgmp) improved its catalytic efficiency at various pH. Furthermore, the Sgmp-treated recombinant LGOX had a subunit structure of α2β2γ2 and nearly the same enzymological character as the LGOX isolated from Streptomyces sp. X-119-6. A higher molecular species observed for the recombinant LGOX was not detected for the Sgmp-treated recombinant LGOX. These results prove that proteolysis by Sgmp is involved in the stabilization of the recombinant LGOX.

Original languageEnglish
Pages (from-to)805-812
Number of pages8
JournalJournal of biochemistry
Volume134
Issue number6
DOIs
Publication statusPublished - Dec 2003

Keywords

  • L-glutamate oxidase
  • Precursor form
  • Proteolysis
  • Streptomyces sp.

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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