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Characterization of novel quinone containing amino acid oxidases from marine bacterium

Research Project

Project/Area Number 17K06926
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Biofunction/Bioprocess
Research InstitutionOkayama University

Principal Investigator

INAGAKI Kenji  岡山大学, 環境生命科学研究科, 教授 (80184711)

Co-Investigator(Kenkyū-buntansha) 今田 勝巳  大阪大学, 理学研究科, 教授 (40346143)
Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords海洋細菌 / Marinomonas mediterranea / グリシンオキシダーゼ / L-リシン ε-オキシダーゼ / バイオセンサー / システイントリプトフィルキノン / 基質特異性 / Marinimonas mediterranea / キノン補酵素 / キノン含有酵素 / アミノ酸オキシダーゼ / X線結晶構造 / 生物工学
Outline of Final Research Achievements

The marine bacterium Marinomonas mediterranea produces L-lysine-ε-oxidase (ε-LysOX) and glycine oxidase (GlyOX), both enzymes are involved in extracellular biofilm formation. ε-LysOX contains a built-in coenzyme, cysteine tryptophyllquinone (CTQ), and is an enzyme with strict substrate specificity. In this study, we succeeded in constructing an E. coli expression system for both enzymes in order to analyze the structures and functions of these enzymes and applied development, and were able to establish a purification system. Furthermore, we clarified that both enzymes have strict substrate specificity and are useful for biosensors, and succeeded in establishing the foundation for industrial application.

Academic Significance and Societal Importance of the Research Achievements

L-リシン-ε-オキシダーゼ(ε-LysOX)は補酵素としてビルトイン型補酵素のシステイントリプトフィルキノン(CTQ)を持つことがアミノ酸オキシダーゼではじめて判明した酵素であり,グリシンオキシダーゼ(GlyOX)も同様にCTQ依存性で,酵素進化的にも極めて興味深い酵素群である。両酵素ともに極めて基質特異性が厳格な酵素で,バイオセンサーを始め,臨床医薬にも貢献が期待される。今本研究の成果により学術的にも産業的にも利用の進展が期待できる。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • 2017 Research-status Report
  • Research Products

    (23 results)

All 2020 2019 2018 2017

All Journal Article (8 results) (of which Peer Reviewed: 6 results,  Open Access: 1 results) Presentation (14 results) Book (1 results)

  • [Journal Article] Marimononas mediterranea由来キノン含有新規グリシンオキシダーゼの大腸菌発現系の構築と性質検討2020

    • Author(s)
      梶山雄輝,溝端佐津紀,赤地周作,根本理子,田村隆,稲垣賢二
    • Journal Title

      岡山大学農学部学術報告

      Volume: 109 Pages: 1-6

    • Related Report
      2019 Annual Research Report
  • [Journal Article] Novel method for L-methionine determination using L-methionine decarboxylase and application of the enzyme for L-homocysteine determination.2020

    • Author(s)
      Okawa, A., Hayashi, M., Inagaki, J., Okajima, T., Tamura, T., and Inagaki, K.
    • Journal Title

      Biosci. Biotechnol. Biochem.

      Volume: 84 Issue: 5 Pages: 927-935

    • DOI

      10.1080/09168451.2020.1715781

    • NAID

      130008122240

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Antiviral effect of sinefungin on in vitro growth of feline herpesvirus type 12019

    • Author(s)
      Kuroda Yudai、Yamagata Haruka、Nemoto Michiko、Inagaki Kenji、Tamura Takashi、Maeda Ken
    • Journal Title

      The Journal of Antibiotics

      Volume: 72 Issue: 12 Pages: 981-985

    • DOI

      10.1038/s41429-019-0234-4

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Application of l-methionine γ-lyase in chiral amino acid analysis2019

    • Author(s)
      Kato, S., Inagaki, K., Oikawa, T.
    • Journal Title

      Anal. Biochem.

      Volume: 580 Pages: 56-61

    • DOI

      10.1016/j.ab.2019.05.018

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Redox-tuning of oxidizing disulfide oxidoreductase generates a potent disulfide isomerase2019

    • Author(s)
      Sutoh Shinya、Uemura Yuko、Yamaguchi Yuko、Kiyotou Asako、Sugihara Rena、Nagayasu Makiko、Kurokawa Mihoko、Ito Koreaki、Tsunekawa Naoki、Nemoto Michiko、Inagaki Kenji、Tamura Takashi
    • Journal Title

      Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

      Volume: 1867 Issue: 3 Pages: 194-201

    • DOI

      10.1016/j.bbapap.2018.12.005

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Redox-tuning of oxidizing disulfide oxidoreductase generates a potent disulfide isomerase.2019

    • Author(s)
      Sutoh,S.,Uemura,Y.,Yamaguchi,Y.,Kiyotou,A.,Sugihara,R.,Nagayasu,M.,Kurokawa,M.,Ito,K.,Tsunekawa,N.,Nemoto,M.,Inagaki,K.,Tamura,T.
    • Journal Title

      Biochim. Biophys. Acta.

      Volume: 1867 Pages: 194-201

    • Related Report
      2018 Research-status Report
  • [Journal Article] The hyperthermophilic cystathionine γ-synthase from the aerobic crenarchaeon Sulfolobus tokodaii: expression, purification, crystallization and structural insights.2017

    • Author(s)
      Sato,D., Shiba,T., Mizuno,S., Kawamura,A., Hanada,S., Yamada,T., Shinozuki,M., Yanagitani,M., Tamura,T., Inagaki,K., and Harada,S.
    • Journal Title

      Acta. Cryst.

      Volume: F73 Pages: 152-158

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Structural and mechanistic insights into homocysteine degradation by a mutant of methionine γ-lyase based on substrate-assisted catalysis.2017

    • Author(s)
      Sato,D., Shiba,T., Yunoto,S., Furutani,K., Fukumoto,M., Kudou,D., Tamura,T., Inagaki,K., and Harada,S.
    • Journal Title

      Protein Science

      Volume: 26 Pages: 1224-1230

    • NAID

      130007628790

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Presentation] 糸状菌由来 L-リシンα-オキシダーゼの活性中心に存在するAsp212残基の機能解析2020

    • Author(s)
      齋藤雅哉,松本侑也,松田崚汰,北川征樹,伊藤菜奈子,今田勝巳,日下部均,根本理子,田村隆,稲垣賢二
    • Organizer
      日本農芸化学会2020年度大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 海洋性細菌由来キノン(CTQ)含有型L-リシン-ε-オキシダーゼの大腸菌発現系構築,精製と性質検討2020

    • Author(s)
      横山大輝,梶山雄輝,赤地周作,根本理子,田村隆,稲垣賢二
    • Organizer
      日本農芸化学会中四国支部第56回講演会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 糸状菌由来 LysOX 二重変異体の基質特異性変換機構2019

    • Author(s)
      北川征樹,松田崚汰,松本侑也,根元理子,田村隆,日下部均,稲垣賢二,今田勝巳
    • Organizer
      酵素・補酵素研究会2019
    • Related Report
      2019 Annual Research Report
  • [Presentation] 糸状菌由来 L-リシンα-オキシダーゼの活性中心に存在するD212残基の機能解析2019

    • Author(s)
      齋藤雅哉,松本侑也,北川征樹,伊藤菜奈子,今田勝巳,日下部均,根本理子,田村隆,稲垣賢二
    • Organizer
      酵素・補酵素研究会2019
    • Related Report
      2019 Annual Research Report
  • [Presentation] 糸状菌L-リシン酸化酵素の前駆体領域による活性制御の構造基盤2019

    • Author(s)
      北川征樹,伊藤菜奈子,松本侑也,天野万里,田村隆,日下部均,稲垣賢二,今田勝巳
    • Organizer
      酵素・補酵素研究会2019
    • Related Report
      2019 Annual Research Report
  • [Presentation] 糸状菌由来LysOX変異体の基質特異性変換の構造基盤2019

    • Author(s)
      北川征樹,松田崚汰,松本侑也,根本理子,田村隆,日下部均,稲垣賢二,今田勝巳
    • Organizer
      第71回日本細胞生物学会大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] L-リシンα-オキシダーゼ変異酵素のX線結晶構造解析と基質認識機構の解析2019

    • Author(s)
      松本侑也,天野万里,齋藤雅哉,北川雄輝,今田勝巳,日下部均,根本理子,田村隆,稲垣賢二
    • Organizer
      日本ビタミン学会第71回大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 高基質特異性酵素L-リシンα-オキシダーゼの基質認識機構の解析2019

    • Author(s)
      松本侑也,天野万里,北川雄輝,今田勝巳,根本理子,日下部均,田村隆,稲垣賢二
    • Organizer
      日本農芸化学会2019年度大会
    • Related Report
      2018 Research-status Report
  • [Presentation] 海洋性細菌Marinomonasu mediterranea由来CTQ依存性グリシンオキシダーゼの精製と性質検討2019

    • Author(s)
      平岩祥太朗,梶山雄輝,溝端佐津紀,赤地周作,根本理子,田村隆,稲垣賢二
    • Organizer
      日本農芸化学会中四国支部第53回講演会
    • Related Report
      2018 Research-status Report
  • [Presentation] 海洋性細菌由来キノン(CTQ)含有型グリシンオキシダーゼの大腸菌発現系の構築及び組換え酵素の精製2018

    • Author(s)
      梶山雄輝,平岩祥太朗,溝端佐津紀,根本理子,田村隆,稲垣賢二
    • Organizer
      第4回日本生物工学会西日本支部講演会
    • Related Report
      2018 Research-status Report
  • [Presentation] 高基質特異性L-アミノ酸オキシダーゼの1種L-リシンα-オキシダーゼから部位特異的変異により作成した新規な芳香族アミノ酸オキシダーゼの性質2018

    • Author(s)
      松本侑也,北川雄輝,今田勝巳,根本理子,田村隆,日下部均,稲垣賢二
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Research-status Report
  • [Presentation] 抗腫瘍性酵素L-リシンα-オキシダーゼの活性中心残基への突然変異による基質認識機構の解析2018

    • Author(s)
      松本侑也,天野万里,北川雄輝,今田勝巳,根本理子,日下部均,田村隆,稲垣賢二
    • Organizer
      日本ビタミン学会第70回大会
    • Related Report
      2018 Research-status Report
  • [Presentation] 高基質特異性L-アミノ酸オキシダーゼは切断され, 高活性で安定になる?2017

    • Author(s)
      松本侑也, 松尾慎作, 天野万里, 根本理子, 田村隆, 伊藤菜奈子, 今田勝己, 日下部均, 稲垣賢二
    • Organizer
      2017年度酵素補酵素研究会
    • Related Report
      2017 Research-status Report
  • [Presentation] 糸状菌由来L-リシンα-オキシダーゼ前駆体の精製, 性質検討およびX線結晶構造解析2017

    • Author(s)
      松本侑也, 天野万里, 伊藤菜奈子, 今田勝己, 根本理子, 日下部均, 田村隆, 稲垣賢二
    • Organizer
      第17回日本蛋白質科学会年会
    • Related Report
      2017 Research-status Report
  • [Book] Methionine Dependence of Cancer and Aging2019

    • Author(s)
      Hoffman, RM., Tan, Y., Li,S., Han, Q., Yagi, S., Takakura, T., Takimoto, A., Inagaki, K., Kudou, D.
    • Total Pages
      25
    • Publisher
      Humana Press (New York)
    • Related Report
      2019 Annual Research Report

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Published: 2017-04-28   Modified: 2021-02-19  

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