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Study on the species-specificity mechanism of C. perfringens lytic enzymes and its application research

Research Project

Project/Area Number 18K07131
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 49050:Bacteriology-related
Research InstitutionMatsuyama University

Principal Investigator

Tamai Eiji  松山大学, 薬学部, 教授 (40333512)

Co-Investigator(Kenkyū-buntansha) 神鳥 成弘  香川大学, 医学部, 教授 (00262246)
成谷 宏文  十文字学園女子大学, 人間生活学部, 教授 (30452668)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywords溶菌酵素 / エンドライシン / オートライシン / ペプチドグリカン / ウエルシュ菌 / ディフィシル菌 / X線結晶構造解析 / 酵素触媒反応機構 / 種特異性 / アミダーゼ / グルコサミニダーゼ / デフィシル菌 / 細胞壁
Outline of Final Research Achievements

Lytic enzymes (endolysins or autolysins) are enzymes that degrade peptidoglycans and kill bacteria. They are classified into four types according to their degradation site: muramidase, glucosaminidase, amidase, and endopeptidase. Furthermore, structurally, they are composed of a catalytic domain and a cell wall binding domains. The aim of this study is to elucidate the substrate recognition and binding mechanism of lytic enzymes at the molecular level. In this study, we clarified the biochemical properties and the structure of the catalytic domain of Psa, an endolysin of C. perfringens. and Acd24020, an autolysin of C. difficile. We found that both of these lytic enzymes are specific to their corresponding bacteria, and that the specificity of Acd24020 depends on the structure of the substrate-binding groove.

Academic Significance and Societal Importance of the Research Achievements

本研究課題では、PsaとAcd24020がそれぞれウエルシュ菌やディフィシル菌に対して特異的に働くことを明らかにし、その特異性が触媒ドメインの構造に依存していることを明らかにした。このことは溶菌酵素の種特異性に関して新たな学術的知見を得ることができた。また、PsaとAcd24020がそれぞれウエルシュ菌とディフィシル菌に特異的に作用し死滅させることを明らかにした。このことは、食中毒やガス壊疽の原因菌であるウエルシュ菌や臨床現場で問題となっている抗菌薬誘発性偽膜生大腸炎の原因菌であるディフィシル菌に対して新たな治療薬の候補となることを明らかにしたものであり、社会的意義が高いと考えられる。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (31 results)

All 2021 2020 2019 2018

All Journal Article (8 results) (of which Peer Reviewed: 8 results,  Open Access: 6 results) Presentation (23 results) (of which Invited: 1 results)

  • [Journal Article] Structural and biochemical characterizations of the novel autolysin Acd24020 from Clostridioides difficile and its full-function catalytic domain as a lytic enzyme.2021

    • Author(s)
      Sekiya, H., Tamai, E., Kawasaki, J., Murakami, K., Kamitori, S.
    • Journal Title

      Mol. Microbiol.

      Volume: 115 Issue: 4 Pages: 684-698

    • DOI

      10.1111/mmi.14636

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] X-ray structures of Clostridium perfringens sortase C with C-terminal cell wall sorting motif of LPST demonstrate role of subsite for substrate-binding and structural variations of catalytic site2021

    • Author(s)
      Tamai Eiji、Sekiya Hiroshi、Nariya Hirofumi、Katayama Seiichi、Kamitori Shigehiro
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 554 Pages: 138-144

    • DOI

      10.1016/j.bbrc.2021.03.106

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A Putative Amidase Endolysin Encoded by Clostridium perfringens St13 Exhibits Specific Lytic Activity and Synergizes with the Muramidase Endolysin Psm2021

    • Author(s)
      Sekiya Hiroshi、Okada Maho、Tamai Eiji、Shimamoto Toshi、Shimamoto Tadashi、Nariya Hirofumi
    • Journal Title

      Antibiotics

      Volume: 10 Issue: 3 Pages: 245-245

    • DOI

      10.3390/antibiotics10030245

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Benzoxazole-Based Metal Complexes to Reverse Multidrug Resistance in Bacteria2020

    • Author(s)
      Kincses Annamaria、Szabo Stefania、Racz Balint、Szemeredi Nikoletta、Watanabe Genki、Saijo Ryosuke、Sekiya Hiroshi、Tamai Eiji、Molnar Joseph、Kawase Masami、Spengler Gabriella
    • Journal Title

      Antibiotics

      Volume: 9 Issue: 10 Pages: 649-649

    • DOI

      10.3390/antibiotics9100649

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Structures of major pilins in Clostridium perfringens demonstrate dynamic conformational change2019

    • Author(s)
      Tamai Eiji、Katayama Seiichi、Sekiya Hiroshi、Nariya Hirofumi、Kamitori Shigehiro
    • Journal Title

      Acta Crystallographica Section D Structural Biology

      Volume: 75 Issue: 8 Pages: 718-732

    • DOI

      10.1107/s2059798319009689

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Synthesis and Antimicrobial Activity of 2-Trifluoroacetonylbenzoxazole Ligands and Their Metal Complexes2018

    • Author(s)
      Watanabe Genki、Sekiya Hiroshi、Tamai Eiji、Saijo Ryosuke、Uno Hidemitsu、Mori Shigeki、Tanaka Toru、Maki Jun、Kawase Masami
    • Journal Title

      Chemical and Pharmaceutical Bulletin

      Volume: 66 Issue: 7 Pages: 732-740

    • DOI

      10.1248/cpb.c18-00158

    • NAID

      130007386740

    • ISSN
      0009-2363, 1347-5223
    • Year and Date
      2018-07-01
    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] Benzoxazole-based Zn(II) and Cu(II) Complexes Overcome Multidrug-resistance in Cancer2018

    • Author(s)
      SPENGLER GABRIELLA、KINCSES ANNAMARIA、RACZ BALINT、VARGA BORISZ、WATANABE GENKI、SAIJO RYOSUKE、SEKIYA HIROSHI、TAMAI EIJI、MAKI JUN、MOLNAR JOSEPH、KAWASE MASAMI
    • Journal Title

      Anticancer Research

      Volume: 38 Issue: 11 Pages: 6181-6187

    • DOI

      10.21873/anticanres.12971

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] Expression of glyceraldehyde-3-phosphate dehydrogenase on the surface of Clostridium perfringens cells.2018

    • Author(s)
      Matsunaga N, Shimizu H, Fujimoto K, Watanabe K, Yamasaki T, Hatano N, Tamai E, Katayama S, Hitsumoto Y.
    • Journal Title

      Anaerobe

      Volume: 51 Pages: 124-130

    • DOI

      10.1016/j.anaerobe.2018.05.001

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] はじめてみようX線結晶構造解析共同研究2 -線毛と溶菌酵素の構造 と機能-2021

    • Author(s)
      玉井栄治
    • Organizer
      第94回細菌学会総会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] ウェルシュ菌由来オートリシンの分子量変化と溶菌活性2021

    • Author(s)
      青野りよ,松永望,玉井栄治,片山誠一,櫃本泰雄
    • Organizer
      第94回細菌学会総会
    • Related Report
      2020 Annual Research Report
  • [Presentation] ウェルシュ菌の溶菌酵素オートリシンの機能解析2021

    • Author(s)
      見尚悟,青野りよ,松永望,成谷宏文,玉井栄治,櫃本泰雄,片山誠一
    • Organizer
      第94回細菌学会総会
    • Related Report
      2020 Annual Research Report
  • [Presentation] ディフィシル菌オートライシンCD24020触媒ドメインのX線結晶解析に基づく基質認識機構の検討2020

    • Author(s)
      神鳥成弘,関谷洋志,玉井栄治
    • Organizer
      第93回日本生化学会大会
    • Related Report
      2020 Annual Research Report
  • [Presentation] ディフィシル菌のエンドペプチダーゼAcd24020の機能解析2020

    • Author(s)
      関谷 洋志, 玉井 栄治
    • Organizer
      第73回日本細菌学会中国・四国支部総会
    • Related Report
      2020 Annual Research Report
  • [Presentation] ウェルシュ菌の自己溶解酵素オートリシンの機能解析2020

    • Author(s)
      江見 尚悟,青野 りよ,松永 望,成谷 宏文,玉井 栄治, 櫃本 泰 雄,片山 誠一
    • Organizer
      第73回日本細菌学会中国・四国支部総会
    • Related Report
      2020 Annual Research Report
  • [Presentation] ウェルシュ菌自己溶菌酵素オートリシンの性状解析2020

    • Author(s)
      青野 りよ, 江草 星良, 松永 望, 玉井 栄治, 片山 誠一, 櫃本 泰雄
    • Organizer
      第73回日本細菌学会中国・四国支部総会
    • Related Report
      2020 Annual Research Report
  • [Presentation] ディフィシル菌のオートリシンCD24020 触媒ドメインの構造と機能解析2020

    • Author(s)
      玉井 栄治,神鳥 成弘,関谷 洋志,河崎 樹里奈,村上 佳穂
    • Organizer
      第93回日本細菌学会総
    • Related Report
      2019 Research-status Report
  • [Presentation] ウェルシュ菌のフィブロネクチン結合タンパク質FbpC, FbpD と溶菌酵素オートリシンの機能解析2020

    • Author(s)
      片山 誠一,江見 尚悟,青野 りよ,河相 朋美,松永 望,玉井 栄治,成谷 宏文,島本 整,櫃本 泰雄
    • Organizer
      第93回日本細菌学会総
    • Related Report
      2019 Research-status Report
  • [Presentation] Detection of fibronectin-binding proteins in membrane vesicle produced by Clostrodium perfringens2020

    • Author(s)
      松永 望,玉井 栄治,片山 誠一,櫃本 泰雄
    • Organizer
      第93回日本細菌学会総
    • Related Report
      2019 Research-status Report
  • [Presentation] ペプチドグリカン上でのウェルシュ菌由来GAPDH とオートリシンの結合2020

    • Author(s)
      青野 りよ,松永 望,玉井 栄治,片山 誠一,櫃本 泰雄
    • Organizer
      第93回日本細菌学会総
    • Related Report
      2019 Research-status Report
  • [Presentation] Biocontrol of Clostridium perfringens by using Two Types of Specific Endolysins2020

    • Author(s)
      成谷 宏文,岡田 真歩,玉井 栄治,関谷 洋志,島本 敏,島本 整
    • Organizer
      第93回日本細菌学会総
    • Related Report
      2019 Research-status Report
  • [Presentation] ウエルシュ菌線毛の構造解析2019

    • Author(s)
      玉井 栄治,神鳥 成弘,関谷 洋志,片山 誠一,成谷 宏文
    • Organizer
      第92回日本細菌学会総会
    • Related Report
      2019 Research-status Report
  • [Presentation] Autolysin and GAPDH as the functioning fibronectin-receptors on the Clostridium perfringens cell surface2019

    • Author(s)
      青野 りよ,藤本 佳那子,松永 望,玉井 栄治,片山 誠一,櫃本 泰雄
    • Organizer
      第92回日本細菌学会総会
    • Related Report
      2019 Research-status Report
  • [Presentation] ディフィシル菌由来グルコサミニダーゼの解析2019

    • Author(s)
      関谷 洋志,西原 剣風,牧 純,玉井 栄治
    • Organizer
      第92回日本細菌学会総会
    • Related Report
      2019 Research-status Report
  • [Presentation] ウエルシュ菌の線毛の構造と機能解析2019

    • Author(s)
      玉井栄治1,片山誠一,関谷洋志,成谷宏文
    • Organizer
      第72回日本細菌学会中国・四国支部総会
    • Related Report
      2019 Research-status Report
  • [Presentation] C. difficile のエンドペプチダーゼCD630_11350 の生化学的解析2019

    • Author(s)
      関谷洋志,玉井栄治
    • Organizer
      第72回日本細菌学会中国・四国支部総会
    • Related Report
      2019 Research-status Report
  • [Presentation] Clostridium perfringens が産生するmembrane vesicle 中のフィブロネクチン結合タンパク質の解析2019

    • Author(s)
      松永 望,玉井栄治,片山誠一,櫃本泰雄
    • Organizer
      第72回日本細菌学会中国・四国支部総会
    • Related Report
      2019 Research-status Report
  • [Presentation] Closutridium perfringens 菌体表層のglyceraldehyde-3-phosphate dehydrogenase(GAPDH)とオートリシンの結合解析2019

    • Author(s)
      青野りよ,松永 望,玉井栄治,片山誠一,櫃本泰雄
    • Organizer
      第72回日本細菌学会中国・四国支部総会
    • Related Report
      2019 Research-status Report
  • [Presentation] ウェルシュ菌の菌体表層に存在するフィブロネクチン(Fn)結合タンパク質(FbpC, FbpD)と溶菌酵素オートリシンの機能解析2019

    • Author(s)
      江見尚悟,青野りよ,河相朋美,松永 望,玉井栄治,成谷宏文,島本 整,櫃本泰雄, 片山誠一
    • Organizer
      第72回日本細菌学会中国・四国支部総会
    • Related Report
      2019 Research-status Report
  • [Presentation] ウェルシュ菌線毛タンパク質CppAとSortase CのX線結晶構造解析2019

    • Author(s)
      神鳥 成弘,玉井 栄治,関谷 洋志,成谷 宏文,片山誠一
    • Organizer
      第92回日本生化学会大会
    • Related Report
      2019 Research-status Report
  • [Presentation] ウェルシュ菌細胞壁分解酵素CPE1138の生化学的解析とX線構造解析2019

    • Author(s)
      玉井 栄治,神鳥 成弘,成谷 宏文,関谷 洋志,渡部 あずさ,牧 純
    • Organizer
      日本薬学会第139年会
    • Related Report
      2018 Research-status Report
  • [Presentation] X-ray structure of Clostridium perfringens CPE1138 catalytic domain hydrolyzing bacterial cell2018

    • Author(s)
      神鳥 成弘,玉井 栄治,関谷 洋志,牧 純,成谷 宏文
    • Organizer
      第91回日本生化学会大会
    • Related Report
      2018 Research-status Report

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Published: 2018-04-23   Modified: 2022-01-27  

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