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MOLECULAR AND GENETICAL STUDIES ON HEMOLYTIC SUBSTANCE PRODUCED BY Pseudomonas cepacia

Research Project

Project/Area Number 03670216
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 細菌学
Research InstitutionYAMAGUCHI UNIVERSITY

Principal Investigator

NAKAZAWA Teruko  YAMAGUCHI UNIVERSITY,SCHOOL OF MEDICINE,PROFESSOR, 医学部, 教授 (40053053)

Co-Investigator(Kenkyū-buntansha) ABE Mitsuko  YAMAGUCHI UNIVERSITY,SCHOOL OF MEDICINE,ASSISTANT, 医学部, 助手 (60107737)
TSUDA Masataka  YAMAGUCHI UNIVERSITY,SCHOOL OF MEDICINE,ASSISTANT PROFESSOR, 医学部, 講師 (90172022)
Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1991: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsPseudomonas cepacia / small hemolytic substance / cepalycin / antifunfal activity / transposon mutagenesis / cepalycin-producing genes / DNA fingerprinting / nosocomial infection / cepalycin 産生遺伝子 / 生理活性物質 / 溶血活性 / 遺伝子クロ-ニング
Research Abstract

1)We established the method for purification of heat-stable hemolytic substance from P.cepacia JN106. Cell-washed supernatant was adsorbed to Amberlite XAD2,eluted with 70% ethanol containing 1mM HCI, and applied on reverse-phase HPLC. Two hemolytic substances,designated cepalycin I and cepalycin II, were isolated. 2)Cepalycins showed the same UV spectra,but the specific activity of cepalycin II was 4 times higher than that of cepalycin I. By LC-FAB Mass analysis,cepalycin II has the molecular weight of 240 with an OH group.Cepalycins were inactivated by alkaline,but resistant to acid.According to the solubility of various solvents,they appeared to be amphipathic molecules. 3)Cepalycins had antifungal activity comparable to amphotericin B. 4)Doughnut-like structures were observed by electron microscopy on a cepalycin-treated membrane of sheep erythrocytes,suggesting the pore-forming activity of the molecule. 5)Four types of non-hemolytic mutants of the strain JN106 were obtained by transposon mutagenesis. DNA fragments of JN106 were cloned that conferred the hemolytic activity to the mutants. 6)Distribution of cepalycin-producing genes were screened among the clinical and environmental isolates of P.cepacia by using the DNA fragment adjacent to the transposon of each mutant as a probe. One of the genes were found to be specific and exclusively for hemolytic strains. Thus ceplaycin is widely distributed among hemolytic strains of P.cepacia. 7)We have carried out DNA fingerprinting of P.cepacia to analyze a nosocomial outbreak of respiratory infection,and proved that the infection was associated with contaminated nebulizer devices.

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] 阿部 光子、津田 雅孝、中澤 晶子: "P.cepaciaの産生する溶血物質の精製とその生合成遺伝子" 日本細菌学会誌. 46. 183- (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 西口 まゆみ、川瀧 美智子、久保田 小百合、瀧川 圭一、山岸 善文、根ヶ山 清、藤田 次郎、森田 敏子、中澤 晶子: "P.cepacia院内感染症の感染源の追及と対策" 環境感染. 7. 33-37 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Takigawa K,Fujita J,Negayama K,Yamagishi Y,Yamaji Y,Ouchi K,Yamada K,Abe M,Nakazawa T: "Nosocomial outbreak of Pseudomonas cepacia respiratory infection in immunocompromized patients with contaminated nebulizer devices" American Review of Respiratory Diseases.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Abe M,Nakazawa T: "Isolation and characterization of new hemolytic substance,cepalycin,from Pseudomonas cepacia"

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Takigawa K,Fujita J,Negayama K,Yamagushi Y,Yamaji Y,Ouchi K,Yamada K,Abe M,Nakazawa T: "Nosocomial outbreak of Pseudomonas cepacia respiratory infection in immunocompromized patients with contaminated nebulizer devices." American Review of Respiratory Diseases.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Abe M,Nakazawa T.: "Isolation and characterization of new hemolytic substance,cepalycin,from Pseudomonas cepacia."

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 西口 まゆみ、川瀧 美智子、久保田 小百合、瀧川 圭一、山岸 善文、根ヶ山 清、藤田 次郎、森田 敏子、中澤 晶子: "P.cepacia院内感染症の感染源の追及と対策" 環境感染. 7. 33-37 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Takigawa K, Fujita J, Negayama K,Yamagishi Y, Yamaji Y,Ouchi K,Yamada K,Abe M, Nakazawa T: "Nosocomial outbreak of Pseudomonas cepacia respiratory infection in immunocompromized patients with contaminated nebulizer devices" American Review of Respiratory Diseases (投稿中).

    • Related Report
      1992 Annual Research Report
  • [Publications] Abe M, Nakazawa T: "Isolation and characterization of new hemolytic substance,cepalycin, from Pseudomonas cepacia"

    • Related Report
      1992 Annual Research Report
  • [Publications] 阿部 光子,津田 雅孝,中澤 晶子: "Pseudomonas cepaciaの産生する溶血物質の精製とのその生合成遺伝子" 日本細菌学雑誌. 46(1). 183 (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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