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Gross culture system for cyanobacteria & its application for medication

Research Project

Project/Area Number 03557024
Research Category

Grant-in-Aid for Developmental Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 細菌学
Research InstitutionJichi Medical School

Principal Investigator

NAKANO Masayasu  Jichi Medical School, Dept.Microbiol., Professor, 医学部, 教授 (70048958)

Co-Investigator(Kenkyū-buntansha) KIRIKAE Teruo  Jichi Med.Sch., Dept.Microbiol., Instructor, 医学部, 助手 (50192563)
SHIRAI Makoto  Ibaraki Univ., School of Agriculture, Associ.Prof., 農学部, 助教授 (10007792)
TAKI Tatuo  Kitasato University, School of Hygiene, Associ.Prof., 衛生学部, 助教授 (70049097)
SAITO Shinji  Jichi Med.Sch., Dept.Microbiol., Instructor, 医学部, 助手 (50195989)
MATUURA Motohiro  Jichi Med.Sch., Dept.Microbiol., Associate Prof., 医学部, 助教授 (20150089)
Project Period (FY) 1991 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥17,100,000 (Direct Cost: ¥17,100,000)
Fiscal Year 1993: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1992: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1991: ¥11,400,000 (Direct Cost: ¥11,400,000)
KeywordsCyanobacteria / Microcystin / Microcystis aeruginosa / antibacterial substance / culture method / Interleukin-6 / monoclonal antibody / gamma-linolenic acid / gamma-linolenic acid / ミクロキスチス / 凍結保存法 / Microcystin / 腫瘍壊死因子 / ミクロキスチン
Research Abstract

A gross culture system for microcystis (cyanobactera) has been developed. When the cells of Microcystis aeruginosa K-139 strain were inoculated (10% v/v) into the system with 10 litters of CB medium (pH 10.0) and were cultured at 28C。 under the conditions of airing (filter-passed air, 9 liters/min) and lighting (4 directions, cool white light, 10, 000 lux). The pH of medium was maintained between 9 and 10, by being adjusted with 1 N HCl solution during the culture period. The cell-growth reached the maximum at day 7, and the yield of cells was 4.56 g. It was 2.5 times greater than that obtained from a stationary culture. The yield of microcystin extracted from the cell-mass was 27.493 mg, and it was 3 times more than that of the statinary culture.
Monoclonal antibodies against microcystin-LR were prepared from hybridoma cell lines. All the antibodies were reacted with not only microcystin-LR but also other microcystins tested. Microcystin-shock in mice could hardly be protected by a previous administration of these antibodies. The microcystin-shock was relieved to some extent when interleukin (IL)-10 was injected 30 min in advance. The serum-level of IL-6 induced by microcystin was lowered by a previous injection of IL-10.
The cell-extract of Microcystis aeruginosa K-18 strain contained anti-bacterial activity against various kinds of gram-positive and negative bacteria. The active substance was purified. However, it was found to be gamma-linolenic acid.

Report

(4 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • 1991 Annual Research Report
  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] S.Saito他: "Cross-reactivity and neutralizing ability of monoclonal antibodies against microcystins" Microbiology and Immunology. 38(印刷中). (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Sakamoto他: "Characteristics of DNA and multiple rpoD homologs of Microcystis(Synechocystis)strains" International Journal of Systemic Bacteriology. 43. 844-847 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 白井 誠他: "Microcystisシアノバクテリアの毒性" 日本細菌学雑誌. 48. 511-522 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Asayama他: "Gene structure and characterization of rpoD and psbA in cyanobacteria" Research in Photosynthesis. 3. 397-400 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Shirai: "The biotechnology of cyanobacteria" Genetic Engineering Research. 5. 23-28 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Harada他: "Isolation of two toxic heptapeptide microcystins from an axenic strain of Microcystis aeruginosa K-139" Toxicon. 29. 479-489 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Saito, S.et al.: "Cross-reactivity and neutralizing ability of monoclonal antibodies against microcystins" Microbiol.Immunol.38 : (in press). (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Sakamoto, T.et al.: "Characteristics of DNA and multiple rpoD homologs of Microcystis (Synechocystis) strains" Int.J.Syst.Bacteriol.43. 844-847 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Shirai, M.et al.: "Toxicity of Microsystis, Cyanobacteria : A Review (in Japanese)" Japn.J.Bacteriol.48. 511-522 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Asayama, M.et al.: "Gene structure and characterization of rpoD and psbA in cyanobacteria" Research in Photosynthesis. 3. 397-400 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Shirai, M.: "The biotechnology of cyanobacteria" Genetic Engineering Res.5. 23-28 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Harada, K.et al.: "Isolation of two toxic heptapeptide microcystins from an axnic strain of Microcystis aeruginosa, K-139" Toxicon. 29. 479-489 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Shirai, M., et al.: "Toxicity and toxins of natural blooms and isolated strains of Microcystis spp. (cyanobacteria) and improved procedure for purification of cultures" Appl.Environ.Microbiol.57. 1241-1245 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Nakano, Y.et al.: "Neutralization of microcystin shocl in mice by tumor necrosis factor alpha antiserum" Appl.Environ.Microbiol.57. 327-330 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Sakamoto 他: "Characteristics of DNA and multiple rpo O homologs of Microcystis (Synechocystis)Strains" International Journal of Systematic Bacteriology. 43. 844-847 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 白井 誠 他: "Microcystis シアノバクテリアの毒性" 日本細菌学雑誌. 48. 511-522 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] S.Saito 他: "Cross-reactivity and neutraliging ability of monoclonal antibodies against microcystins" Microbiology and Immunology. 38(印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Asayama,M.,et al.: "Gene structure and characterigation of rpoD and psbA in cyanobacteria" Current research in photosynthesis (Kluwer Academic Publishers).

    • Related Report
      1992 Annual Research Report
  • [Publications] Harada,K.,et al.: "Isolation of two toxic heptapeptide microcystins from an axenic strain of Microcystis aeruginosa,K-139" Toxicon. 29. 479-489 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Shirai,M.,et al.: "Toxicity and toxins of natural blooms and isolated strains of Microcystis spp.(Cyanobacteria)and improved procedure for purification of cultures" Appl.Environ.Microbiol.57. 1241-1245 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Nakano,Y.,et al.: "Neutralization of microcystin shock in mice by tumor necrosis factor alpha antiserum" Appl.Enriron.Microbiol.57. 327-330 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Nakano,Y.et al.: "Neutralization of microcystin shock in mice by tumor necrosis factor alpha antiserum." Appl.Environ.Microbiol.57. 327-330 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Shirai,M.et al.: "Toxicity and toxins of natural blooms and isolated strains Microcystis spp.(Cyanobacteria)and improved procedure for purification of cultures." Appl.Environ.Microbiol.57. 1241-1245 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Harada,K.et al.: "Isolation of two toxic heptapeptide microcystins from an axenic strain of Microcystis aeruginosa,K-139." Toxicon. 29. 479-489 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Shirai,M.et al.: "Mass culture of unicellular cyanobacteria,Microcystis spp." Appl.Enviroe.Microbiol.

    • Related Report
      1991 Annual Research Report
  • [Publications] Shirai,M.et al.: "Release of microcystin from Microcystis cells." Appl.Environ.Microbiol.

    • Related Report
      1991 Annual Research Report

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

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