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Development of mutacin MT6223 from Streptococcus sobrinus as an anti-caries agent

Research Project

Project/Area Number 06557099
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section試験
Research Field 病態科学系歯学(含放射線系歯学)
Research InstitutionOsaka University

Principal Investigator

HAMADA Shigeyuki  Osaka University, Faculty of Dentistry, Professor, 歯学部, 教授 (60028777)

Co-Investigator(Kenkyū-buntansha) TANAKA Takaharu  Suntory Institute for Biomedical Research, Director, 第一創薬研究所, 所長
OOSHIMA Takashi  Osaka University, Faculty of Dentistry, Associate Professor, 歯学部, 助教授 (80116003)
FUJIWARA Taku  Osaka University, Faculty of Dentistry, Assistant Professor, 歯学部・附属病院, 講師 (00228975)
高橋 一郎  大阪大学, 歯学部, 助手 (20206791)
Project Period (FY) 1994 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥9,300,000 (Direct Cost: ¥9,300,000)
Fiscal Year 1996: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1995: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1994: ¥3,600,000 (Direct Cost: ¥3,600,000)
Keywordsmutacin / Streptococcus mutans / dental caries / Streptococcus sobrinas / bacteriocin / う蝕
Research Abstract

Streptococcus sobrinus MT6223 extracellularly produces mutacin, a bactericidal agent. We have attempted to purify and characterize this bacteriocin.
1.Dialyzed TTY (dTTY) broth was better than BHI or Eagle medium for the growth of Streptococcus sobrinus MT6223. Production of mutacin MT6223 was also the highest among the three medium when cultured in dTTY.
2.After culture supernatant from dTTY-grown S.sobrinus was precipitated with ammonium sulfate, the resultant precipitant was applied onto several column chromatography. Recovery of mutacin using Sepharose column was more efficient than Bio-Gel or Sephadex.
3.Mutacin was purified using Sepharose CL-6B,Develosil ODS-HG-5, and Resource RPC column chromatography. It was found that the bacteriocin is heat and acid-stable, and that mutacin MT6223 is composed of peptide. Since mutacin in 1 M urea solution exerted killing bactericidal activity against S.mutans MT8148, the bacteriocin proved to exhibit the anti-microbial activity as a monomeric form.
4.FAB/MS and MALDI-TOF MS analyzes revealed that mutacin possesses a molecular mass of 2,716. Amino acid analysis determined that the N-terminal sequence is Ala-Val-X-and composition of mutacin is rich with Gly and Ala.
5.We have isolated a variant of S.sobrinus that can produce mutacin by fourfold higher compared with parent strain MT6223. Purification of mutacin from this high producer is now in progress using the above methods established in this project.

Report

(4 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] T. Ooshima: "Detection of caries-inducing microorganisms in hyposalivated rats without infection of mutans streptococci" Microbiology and Immunology. 38・1. 39-45 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T. Ooshima: "Reduction of dental plaque deposition in human by oolong tea extract" Caries Research. 28. 146-149 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T. Fujiwara: "Deletion and reintroduction of glucosyltransferase genes of streptococcus mutans and role of their gene product in sucrose dependent cellular adherence" Microbial Pathogenesis. 20. 225-233 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 浜田茂幸: "ミュータンスレンサ球菌のビルレンス因子の解析と同因子の抑制" 日本細菌学雑誌. 51・4. 931-951 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] S. Hamada: "Peroxidase-catalyzed generation of catechin oligomers that inhibit glucosyltransferases from Streptococcus sobrinus" FEMS Microbiology Letters. 143. 35-40 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Ooshima et al.: "Detection of caries-inducing microorganisms in hyposalivated rats without infection of mutans streptococci" Microbiology and Immunology. 38(1). 39-45 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Ooshima et al.: "Reduction of dental plaque deposition in humans by oolong tea extract" Caries Research. 28. 146-149 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Fuiwara et al.: "Delection and reintroduction of glucosyltransferase genes of Streptococcus mutans and role of their gene products in sucrose dependent cellular adherence" Microbial Pathogenesis. 20. 225-233 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] S.Hamada et al.: "Peroxidase-catalyzed generation of catechin oligomers that inhibit glucosyltransferases from Streptococcus sobrinus." FEMS Microbiology Letters. 143. 35-40 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] S.Hamada: "Peroxidase-catalyzed generation of catechin oligomers that inhibit glucosyltransferase from Streptococcus sobrinus" FEMS Microbiology Letters. 143. 35-40 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Fujiwara: "Deletion and reintroduction of glucosyltransferase genes of Streptococcus mutans and role of their gene products in sucrose dependent cellular adherence" Microbial Pathogenesis. 20. 225-233 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Fujiwara: "Deletion and reintroduction of glucosyttunferase gones of S mutans and sole of then gene products insuibl dependent cellularadference" Microbial Pathogenesis. 20(印刷中). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hamada,S.: "Development of preventive measures based on the aetidogy of dental caries" Microbial Ecology in Health and Disease. 8(in press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] H.Shibata: "Molecular characterization of an extracellular bacteriocin from S.sobrinus" Abstracts of the 95th General Meeting of the ASM. (in press). (1995)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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