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Development of new hypertermophiles and utilization of their products

Research Project

Project/Area Number 05556034
Research Category

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

Allocation TypeSingle-year Grants
Research Field General fisheries
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

ISHIDA Yuzaburo  Kyoto Univ., Agriculture, Professor, 農学部, 教授 (20026488)

Co-Investigator(Kenkyū-buntansha) ISHINO Yoshizumi  Takara Shuzo, Biotechnology Res.Lab., Senior Researcher, 中央研究所, 主任研究員
OWADA Koichi  Univ.Tokyo, Inst.Oceanography, Professor, 海洋研究所, 教授 (30013585)
SAKO Yoshihiko  Kyoto Univ., Agirulture, Lecturer, 農学部, 講師 (60153970)
UCHIDA Aritsune  Kyoto Univ., Agriculture, Associate Prpfessor, 農学部, 助教授 (50027190)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥5,500,000 (Direct Cost: ¥5,500,000)
Fiscal Year 1994: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1993: ¥4,000,000 (Direct Cost: ¥4,000,000)
KeywordsHyperthermophile / Heat stable enzyme / Hydrothermal vent / Marine bacteria / Archaea / Protease / DNAポリメラーゼ / 深海
Research Abstract

Recent isolations and descriptions of new microorganisms from the hydrothermal vents(>200゚C)have been of ecological, physiological, and biotechnological interest, especially with respect of hyperthermophile. Its has not made use of these hyperthermohpiles because of difficulty in cultivation under hightemperature and pressure. In this study, we firstly establish the isolation and cultivation system of marine hyperthemophiles growing at 80 to 100゚C which are available for biotechnology and then attempt to use heat stable enzymes for biotechnological purpose. The results are as follows ;
(1)The anaerobic cultivation system is developed by use of heat and high pressure tolerant glass bottlea and butyl rubber stoppers containing a gas mixture of 80%H_2 and 20%CO_2 at 0.2MPa. For culturing aerobic organisms at temperature up to 95゚C the new gyrotary shaker is developed by composing of heat torelant grease and super insulating materials.
(2)We isolated new anaerobic hypethermophiles Pyrococcus TS100, Methanococcus TS300, aerobic Protothermus maritima OKD7 from shallow hydrothermal vents in Tachibana bay, Nagasaki and the novel marine hyperthermophilic Archaon Aeropyrus pernix Kl from Kodakara island, Kagoshima. Optimum temperature of TS100, TS300, OKD7and Kl are 103,88,80and90-95゚C,respectively. The sequences of 16SrDNA shows that A.pernix Kl in a primitive Crenarchaeta and P.maritima OKD7 is a primive the Bacteria.
(3)Nonallosteric CO_2 fixing enzyme (phosphoenolpyruvate carboxylase) was purified and characterized from methanogenic bacterium TS300, therefore it might be useful for diagnosis of cardiac insufficiency. A.pernix Kl produces a useful extracellular metalloproteinase which shows optimum temperature of 110゚C and theextreme themostability (half life at 120゚C,150min). DNA polymerase of Pyrococcus TS100 is a very thermostable and useful for PCR method.

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] T.Hoaki et al.: "Amino acid requirements of two hyperthermophilic archaeral isolates from deep sea vents:Desulfurococcus strain SY and Pyrococcus strain GB-D." Appl.Env.Microbiol.59. 610-613 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Hoaki et al.: "Growthrequirements of hyperthermophilic sulfur-dependent heterotrophic archaea isolated from a shallow submarine geothermal system with reference to their essential amino acids." Appl.Env.Microbiol.60. 2898-2904 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Uemori et al.: "Cloning of the Bacillus caldotenax DNA polymerase gene and characterization of the gene product." J.Biochem.113. 401-410 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Ishino et al.: "Amino acid sequence motifs essential to 3'→5'exonuclease activity of Escherichia coli DNA polymerse II." J.Biol.Chem.269. 14655-14660 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Ishino et al.: "Overproduction of Thermusaquaticus DNA polymerase and its structural analysis by ion-spray mass spectrometry." J.Biochem.(in press). (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Sako et al.: "Aeropyrus pernix gen nov.,sp.nov.:an Aerobic,Marine hyperthemohpilic Archeon" Int.J.Syst.Bacteriol.(投稿中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 左子芳彦: "二酸化炭素-化学・生化学・環境-" 東京化学同人, 192 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 向井博之, 石野良純: "PCR法-分子生物学に及ぼすその革命的影響" 東京化学同人, 70 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Hoaki et al.: "Amino acid requirements of two hyperthermophilic archaeral isolates from deep sea vents : Desulfurococcus strain SY and Pyrococcus strain GB-D." Appl.Env.Microbiol.59. 610-613 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Hoaki et al.: "Growth requirements of hyperthermophilic sulfurdependent heterotrophic archaea isolated from a shallow submarine geothermal system with reference to their essential amino acids." Appl.Env.Microbiol.60. 2898-2904 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Uemori et al.: "Cloning of the Bacillus caldotenax DNA polymerase gene and characterization of the gene product." J.Biochem.113. 401-410 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Ishino et al.: "Amino acid sequence motifs essential to 3'*5' exonuclease activity of Escherichia coli DNA polymerse II." J.Biol.Chem.269. 14655-14660 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Ishino et al.: "Overproduction of Thermus aquaticus DNA polymerase and its structural analysis by ion-spray mass spectrometry." J.Biochem.in press.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Sako et al.: "Restriction fragment length polymorphism of ribosomal DNA internal transcribed spacer and 5.8S regions in Japanese Alexandrium species (Dinophyceae)." J.Phycol.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Sako: CO_2-Chemistry, Biochemistry, Environment-. Tokyo Kagaku Dojin, 192 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] H.Mukai: PCR method-Evolutional effect to molecular biology. Tokyo Kagaku Dojin, in press, (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Hoaki et al.: "Amino acid requirements of two hyperthermophilic archaeral isolates from deep sea vents:Desulfurococcus strain SY and Pyrococcus strain GB-D." Appl.Env.Microbiol.59. 610-613 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Hoaki et al.: "Growth requirements of hyperthermophilic sulfur-dependent heterotrophic archaea isolated trom a shallow submarine geothermal system with reterence to their essential amino acids." Appl.Env.Microbiol.60. 2898-2904 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Uemori et al.: "Cloning of the Bacillus caldotenax DNA polymerase gene and characterization of the geneproduct." J.Biochem.113. 401-410 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] Y.Ishino et al.: "Amino acid sequence motifs essential to 3'→5'exonuclease activity of Escherichia coli DNA polymerse II." J.Biol.Chem.269. 14655-14660 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Y.Ishino et al.: "Overproduction of Thermus aquaticus DNA polymerase and its structural analysis by ion-spray mass spectrometry." J.Biochem.in press

    • Related Report
      1994 Annual Research Report
  • [Publications] M.Adachi et al.: "Restriction fragment length polymorphism of ribosomal DNA internal transcribed spacer and 5.8S regions in Japanese Alexandrium species(Dinophyceae)." J.Phycol.30. 857-863 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 左子 芳彦: "二酸化炭素-化学・生化学・環境-" 東京化学同人, 192 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 向井 博之、石野 良純: "PCR法-分子生物学に及ぼすその革命的影響" 東京化学同人, 70 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] A.Uchida: "Incorporation of methionine into dimethylthiopropanoic acid in the dinoflagellate Crypthecodinium cohnii." Nippon Suisan Gakkaishi. 59. 851-855 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] T.Mitsuhashi: "Effect of organic acids on the swelling of Bacillus subtilis spores during germination." Nippon Suisan Gakkaishi. 59. 847-850 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Adachi: "Application of monoclonal antibodies to field samples of Alexandrium species." Nippon Suisan Gakkaishi. 59. 1171-1175 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Shrestha: "A note on isozyme analysis of F1 progeny of Gonium pectorale" Bull.Jap.Soc.Micro.Ecol.8. 43-46 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Adachi: "Cross-reactivity of five monoclonal antibodies to various isolates of Alexandrium as determined by an indirect immunofluorescence method" Nippon Suisan Gakkaishi. 59. 1307 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 左子芳彦: "海洋特殊環境の微生物-超好熱細菌-" バイオサイエンスとインダストリー. (印刷中). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 左子芳彦: "「二酸化炭素:化学、生化学、環境」" 東京化学同人(印刷中), (1994)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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