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

• Arima Jiro

  Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University

• Tamura Takashi

  Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University

• Kusakabe Hitoshi

  Research Laboratories, Yamasa Shoyu Co., Ltd.

• Ashiuchi Makoto

  Department of Bioresources Science, Faculty of Agriculture, Kochi University

• Yagi Toshiharu

  Department of Bioresources Science, Faculty of Agriculture, Kochi University

• Tanaka Hidehiko

  Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University

• Inagaki Kenji

  Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University

書誌事項

タイトル別名

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

抄録

L-Glutamate oxidase (LGOX) from Streptomyces sp. X-119-6 is a protein of 150 kDa that has hexamer structure α₂β₂γ₂. 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.

収録刊行物

• The Journal of Biochemistry

  The Journal of Biochemistry 134 (6), 805-812, 2003

  社団法人 日本生化学会

キーワード

• L-glutamate oxidase
• precursor form
• proteolysis
• <i>treptomyces</i> sp.

詳細情報

• CRID: 1390001204964836224
• NII論文ID: 130003534631
• NII論文ID: 10012059658
• NII書誌ID: AA00694073
• DOI: 10.1093/jb/mvg206
• COI: 1:CAS:528:DC%2BD2cXis1Kksrg%3D
• ISSN: 17562651
• ISSN: 0021924X
• NDL書誌ID: 6830681
• PubMed: 14769868
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I6830681
• Web Site: http://academic.oup.com/jb/article-pdf/134/6/805/2380750/mvg206.pdf
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可