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Development of new antibiotics targeting cell wall biosynthesis of oral pathogenic bacteria

Research Project

Project/Area Number 12557186
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 矯正・小児・社会系歯学
Research InstitutionKyushu University (2003)
Nihon University (2000-2002)

Principal Investigator

YAMASHITA Yoshihisa  Kyushu University, Faculty of Dental Science, Professor, 歯学研究院, 教授 (20192403)

Co-Investigator(Kenkyū-buntansha) NOKANO Yoshio  Kyushu University, Faculty of Dental Science, Associate Professor, 歯学研究院, 助教授 (80253459)
高橋 富久  日本大学, 歯学部, 助教授 (40246905)
Project Period (FY) 2000 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥11,900,000 (Direct Cost: ¥11,900,000)
Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2002: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2001: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2000: ¥4,500,000 (Direct Cost: ¥4,500,000)
KeywordsStreptococcus mutans / Actinobacillus actinomycetemcomitans / cell wall polysaccharide / antibiotics / S.mutans / A.actinomycetemcomitans / 莢膜様多糖 / ABCトランスポーター / 齲蝕細菌 / 歯周病細菌 / 薬剤耐性 / 口腔病原性細菌 / 抗生剤 / Actinobacillus actinomycetemcomitans / Streptococcus mutans
Research Abstract

(The 2000 fiscal year)The genes which were related to the synthesis of the capsule polysaccharide of A.actinomycetemconutans(Aa) and cell wall polysaccharide of S.mutans was cloned. Homologous research for the determined sequence of the cloned genes revealed that some of them seemed to encode glycosyltransferases.
(The 2001 fiscal year)The detailed function of each gene product was analyzed with the expressed gene products in Escherichia coli. The result indicated that the gene product of rgpA which is located in the uppermost stream of rgp gene cluster was glycosyltransferase which catalyzes the transfer of the first rhamnose residue. Furthermore, it became to be obvious that the serotype specific cell wall polysaccharide was important for the drug tolerance of S.mutans by comparing the drug tolerance for antibiotics of wild type strain Xc and its mutant strain Xc41.
(The 2002 fiscal year)The genes which encode GDP-D-mannose 4,6-dehydratase and GDP-keto-6-deoxy-D-mannose reductase were … More cloned from the chromosome of the serotype a Aa SUNYaB75 strain. In addition, construction of the variant mutant strains Aa which lost serotype a,c, and d capsular polysaccharide antigens was succeeded by the insertional inactivation. When the tolerance of these variants for several antimicrobial drugs was examined, the sensitivity for bacitracin and vancomycin also slightly increased in any variants.
(The 2003 fiscal year)The antibodies were raised against the rgpC and rgpD gene products of S.mutans expressed in and purified from E.coli. Though S.mutans was cultivated in the presence of the obtained antibodies, it was not possible to effectively inhibit the cell wall polysaccharide synthesis. When the antibody was taken in liposome and added to the culture medium, the interference tendency in the cell wall polysaccharide synthesis was slightly recognized. In case of Aa, antibodies against ABC transporters concerning the synthesis of the polysaccharide were prepared. Every antibodies showed slight blocked effect on capusular polysaccharide synthesis. Less

Report

(5 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] Shibata, Y., Yamashita, Y., Ozaki, K., et al.: "Expression and characterization of streptococcal rgp genes required for rhamnan synthesis in Escherichia coli"Infect.Immun.. 70. 2891-2898 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tsuda, H., Yamashita, Y., Shibata, Y., et al.: "Genes involved in bacitracin resistance in Streptococcus mutans"Antimicrob.Agents Chemother. 46. 3756-3764 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Suzuki, N., Nakano, Y., Yoshida, Y., et al.: "Guanosine diphosphate-4-keto-6-deoxy-D-mannose reductase in the pathway for the synthesis of GDP-6-deoxy-D-talose in Actinobacillus actinomycetemcomitans"Eur.J.Biochem.. 269. 5963-5971 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ozaki, K., Shibata, Y., Yamashita, Y., et al.: "A novel mechanism for glucose side-chain formation in rhamnose-glucose polysaccharide synthesis"FEBS Letters. 532. 159-163 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Engels-Deutsch, M., Pini, A., Yamashita, Y., et al.: "Insertional inactivation of pac and rmlB genes reduces the release of tumor necrosis factor alpha, interleukin-6, and interleukin-8 induced by Streptococcus mutans in monocytic, dental pulp, and periodontal ligament cells"Infect.Immun.. 71. 5169-5177 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shibata, Y., Ozaki, K., Seki, M., et al.: "Analysis of loci required for determination of serotype antigenicity in Streptococcus mutans and its clinical utilization"J.Clin.Microbiol.. 4107-4112 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shibata, Y., Yamashita, Y., Ozaki, K., Nakano, Y., Koga, T.: "Expression and characterization of streptococcal rgp genes required for rhamnan synthesis in Escherichia coli"Infect.Immun.. 70. 2891-2898 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Tsuda, H., Yamashita, Y., Shibata, Y., Nakano, Y., Koga, T.: "Genes involved in bacitracin resistance in Streptococcus mutans."Antimicrob.Agents Chemother.. 46. 3756-3764 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Suzuki, N., Nakano, Y., Yoshida, Y., Nezu, T., Terada, Y., Yamashita, Y., Koga, T.: "Guanosine diphosphate-4-keto-6-deoxy-D-mannose reductase in the pathway for the synthesis of GDP-6-deoxy-D-talose in Actinobacillus actinomycetemcomitans."Eur.J.Biochem.. 269. 5963-5971 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ozaki, K., Shibata, Y., Yamashita, Y., Yamashita, Y., Tsuda, H., Koga, T.: "A novel mechanism for glucose side-chain formation in rhamnose-glucose polysaccharide synthesis."FEBS Letters. 532. 159-163 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Engels-Deutsch, M., Pini, A., Yamashita, Y., Shibata, Y., Haikel, Y., Scholler-Guinard, M., Klein, J.P.: "Insertional inactivation of pac and rmlB genes reduces the release of tumor necrosis factor alpha, interleukin-6,and interieukin-8 induced by Streptococcus mutans in monocytic, dental pulp, and periodontal ligament cells."Infect.Immun.. 71. 5169-5177 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shibata, Y., Ozaki, K., Seki, M., Kawato, K., Tanaka, H., Nakano, Y., Yamashita, Y.: "Analysis of loci required for determination of serotype antigenicity in Streptococcus mutans and its clinical utilization."J.Clin.Microbiol.. 41. 4107-4112 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shibata, Y. et al.: "Analysis of loci required for determination of serotype antigenicity in Streptococcus mutans and its clinical utilization"Journal of Clinical Microbiology. 41. 4107-4112 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Engels-Deutsch, M. et al.: "Insertional inactivation of pac and rmlB genes reduces the release of tumor necrosis factor alpha, interleukin-6, and interleukin-8 induced by Stretococcus-----"Infection and Immunity. 71. 5169-5177 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Shibata, Y., Yamashita, Y, Oaki, K., Nakano, Y., Koga, T.: "Expression and characterization of streptococcal rgp gene required for rhamnan synthesis in Escherichia coli"Infection and Immunity. 70・6. 2891-2898 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Tsuda, H., Yamashita, Y., Shibata, Y., Nakano, Y., Koga, T.: "Genes involved in bacitracin resistance in Streptococcus mutans"Antimicrobial Agents and Chemotherapy. 46・12. 3756-3764 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Suzuki, N., Nakano, Y, Yoshida, Y, Nezu, T., Terada, Y., Yamashita, Y, Koga, T.: "Guanosine diphosphate-4-keto-6-deoxy-D-mannose reductase in the pathway for the synthesis of GDP-6-deoxy-D-talose in Actinobacillus actinomycetemcomitans"European Journal of Biochemistry. 269・23. 5963-5971 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Oaki, K., Shibata, Y., Yamashita, Y, Nakano, Y., Tsuda, H., Koga, T.: "A novel mechanism for glucose side-chain formation in rhamnose-glucose polysaccharide synthesis"FEBS Letters. 532. 159-163 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Shibata, Y., Yamashita, Y.et al.: "Characterization of the rgp genes required for rhamnan synthesis in streptococci"Infection and Immunity. (In press).

    • Related Report
      2001 Annual Research Report
  • [Publications] Y.Yoshida et al.: "A gene cluster for the synthesis of serotype d-specific polysaccharide-."Biochimica et Biophysical Acta. 1493巻. 259-263 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Y.Nakano et al.: "Thymidine diphosphate-6-deoxy-L-lyxo-4-hexulose reductase synthesis-."Journal of Biological Chemistry. 275巻10号. 6806-6812 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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