A mechanism for regulation of clostripain activity and modification of inflammatory response by the enzyme.

2011 Fiscal Year Final Research Report

• Project/Area Number: 21590483
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bacteriology (including Mycology)
• Research Institution: Chugoku Gakuen University (2011); Kagawa University (2009-2010)
• Principal Investigator: OKABE Akinobu 中国学園大学, 現代生活学部, 教授 (20093677)
• Co-Investigator(Kenkyū-buntansha): MIYATA Shigeru 中部大学, 応用生物学部, 准教授 (90314913) ; NARIYA Hirofumi 香川大学, 医学部, 助教 (30452668) ; SUZUKI Motoo 香川大学, 医学部, 助教 (80457340)
• Project Period (FY): 2009 – 2011
• Keywords: ウェルシュ菌 / clostripain / プロテアーゼ / 病原因子 / 凝集性

Research Abstract

To examine a pathogenic role of clostripain(Clp) produced by Clostridium perfringens, Clp mutant and wild-type strains were injected into the mouse femur. Both strains showed gas gangrene and leukostasis to a similar extent, but no significant difference in histological characteristics. We developed an in-frame deletion method, by which we constructed various mutant strains. To obtain various toxins and mutant enzymes in large quantities, we constructed xylose-inducible expression system. Although we attempted to analyze regulation of Clp activity by a linker peptide and also an effect of Clp on eat-me signal of polymorphonuclear leukocytes, we failed to obtain conclusive evidence. We found the Clp mutant aggregates upon heating. Addition of EDTA, an inhibitor of Clp, to the culture of the wild type strain caused self-aggregation.

Research Products

• [Journal Article] Development and characterization of a xylose-inducible gene expression system for Clostridium perfringens (2011)
  • Author(s): Hirofumi Nariya, Shigeru Miyata, Tomomi Kuwahara, and Akinobu Okabe
  • Journal Title: Applied and Environmental Microbiology Volume: 77-23 Pages: 8439-8441
  • Peer Reviewed
• [Journal Article] Identification and characterization of a putative endolysin encoded by episomal phage phiSM101 of Clostridium perfringens (2011)
  • Author(s): Hirofumi Nariya, Shigeru Miyata, Eiji Tamai, Hiroshi Sekiya, Jun Maki, and Akinobu Okabe
  • Journal Title: Applied Microbiology and Biotechnology Volume: 90-6 Pages: 1973-1979
  • Peer Reviewed
• [Journal Article] Development and application of a method for counterselectable in-frame deletion in Clostridium perfringens (2011)
  • Author(s): Hirofumi Nariya, Shigeru Miyata, Motoo Suzuki, Eiji Tamai, and Akinobu Okabe
  • Journal Title: Applied and Environmental Microbiology Volume: 77-4 Pages: 1375-1382
  • Peer Reviewed
• [Journal Article] High-level production and purification of clostripain expressed in a virulence-attenuated strain of Clostridium perfringens (2011)
  • Author(s): Hiroaki Tanaka, Hirofumi Nariya, Motoo Suzuki, Hitoshi Houchi, Eiji Tamai, Shigeru Miyata, and Akinobu Okabe
  • Journal Title: Protein Expression and Purification Volume: 76-1 Pages: 83-89
  • Peer Reviewed
• [Journal Article] Purification and characterization of a clostripain-like protease from a recombinant Clostridium perfringens culture (2010)
  • Author(s): Sadao Manabe, Hirofumi Nariya, Shigeru Miyata, Hiroaki Tanaka, Junzaburo Minami, Motoo Suzuki, Yuki Taniguchi, and Akinobu Okabe
  • Journal Title: Microbiology Volume: 156-2 Pages: 561-569
  • Peer Reviewed
• [Presentation] Development and characterization of a xylose-inducible gene expression system for Clostridium perfringens (2011)
  • Author(s): Hirofumi Nariya, Shigeru Miyata, and Akinobu Okabe
  • Organizer: XIII International Congress of Bacteriology and Applied Microbiology, International Union of Microbiological Societies Congresses 2011
  • Place of Presentation: Sapporo, Japan
  • Year and Date: 20110906-16
• [Presentation] High-level production and purification of clostripain expressed in a virulence-attenuated strain of Clostridium perfringens (2011)
  • Author(s): Hiroaki Tanaka, Hirofumi Nariya, Motoo Suzuki, Shigeru Miyata, Eiji Tamai, Shinji Kosaka, Noriyasu Fukuoka, Hitoshi Houchi, and Akinobu Okabe
  • Organizer: XIII International Congress of Bacteriology and Applied Microbiology, International Union of Microbiological Societies Congresses 2011
  • Place of Presentation: Sapporo, Japan
  • Year and Date: 20110906-16
• [Presentation] ウェルシュ菌のglyceraldehyde-3-phosphate dehydrogenaseの性状解析 (2010)
  • Author(s): 田中裕章, 成谷宏文, 鈴木基生, 谷口有紀, 芳地一, 宮田茂, 岡部昭延
  • Organizer: 第63回日本細菌学会中国・四国支部総会
  • Place of Presentation: 松山
  • Year and Date: 2010-10-16
• [Remarks] 報告書等(1)岡部昭延,成谷宏文,宮田茂."ウェルシュ菌のClostripain-like proteaseの病原激意義についての研究"乳酸菌研究会に関する報告書平成21年度300-303, 2010.
• [Remarks] (2)岡部昭延,宮田茂,成谷宏文."Clostridium perfringensの菌体表面のglyceraldehyde-3-phosphate dehydrogenaseの研究"乳酸菌研究会に関する報告書平成22年度274-277, 2011.
• [Remarks] (3)坂本知世,杉井美香,平田知之;担当教員岡部昭延."ウェルシュ菌の自己凝集に関する研究"平成23年度卒業研究中国学園大学現代生活学部,人間栄養学科7期生印刷中. 2012.
• [Remarks]
  • URL: http://www.med.kagawa-u.ac.jp/faculty/center/igaku_kouza/bunshi_biseibutsu/
• [Remarks]
  • URL: http://www.cjc.ac.jp/eiyo/index.html