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2005 Fiscal Year Final Research Report Summary

Experimental test of the hyperthermophilic common ancestor hypothesis.

Research Project

Project/Area Number 16570189
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Evolutionary biology
Research InstitutionTokyo University of Pharmacy and Life Sciences

Principal Investigator

YAMAGISHI Akihiko  Tokyo Univ.Pharm.Life Sci, Sch.Life Sci., Prof., 生命科学部, 教授 (50158086)

Project Period (FY) 2004 – 2005
Keywordscommon ancestor / hyperthermophile / Archaea / Eubacteria / Thermostability of enzyme / isopropylmalate dehydrogenase
Research Abstract

In order to test experimentally the hypothesis that the common ancestor of all the living organisms on the earth was hyperthermophilic, we designed and conducted a series of experiments. We constructed the phylogenetic tree of the two closely related enzymes : isopropylmalate dehydrogenase (IPMDH) and isocitrate dehydrogenase. The ancestral sequence of these enzymes possessed by the common ancestor of all the living organisms was inferred using the phylogenetic tree. Each ancestral amino acid residue was individually introduced to the IPMDH of an extreme thermophile Thermus thermophilus as a mutation. The mutant IPMDHs was expressed in Escherichia coli and purified. The thermostability of each ancestral mutant IPMDH was estimated. The 6 out of 12 ancestral mutants showed hither thermal stability than the original T.thermophilus IPMDH. However, there still remains the possibility that the effect of ancestral mutation resulted from the fact that the ancestral residue is conserved. However, the statistic analysis showed that the stabilization effect can be attributed to the antiquity of the residues but not to the conservation of the residues.
The multiple ancestral mutant IPMDHs each having multiple ancestral mutant residues were constructed and analyzed. The multiple ancestral mutant IPMDHs showed higher stability than single point ancestral mutant IPMDHs. The multiple and single-point ancestral mutant IPMDHs have simillar or even improved enzymatic activity.
The ancestral NDT having ancestral residues were constructed and showed high thermal stability.

  • Research Products

    (8 results)

All 2006 2005 2004

All Journal Article (7 results) Book (1 results)

  • [Journal Article] Designing thermostable proteins : Ancestral mutants of 3-isopropylmalate dehydrogenase designed by using a phylogenetic tree2006

    • Author(s)
      K.Watanabe, T.ohkuri, S.Yokobori, A.Yamagishi
    • Journal Title

      J. Mol. Biol., 355

      Pages: 664-674

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Designing thermostable proteins : Ancestral mutants of 3-isopropylmalate dehydrogenase designed by using a phylogenetic tree2006

    • Author(s)
      K.Watanabe, T.ohkuri, S.Yokobori, A.Yamagishi
    • Journal Title

      J.Mol.Biol. 355

      Pages: 664-674

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Thermostability of ancestral mutants of Caldococcus noboribetus isocitrate dehydrogenase2005

    • Author(s)
      H.Iwabata, K.Watanabe, T.Ohkuri, S.Yokobori, A.Yamagishi
    • Journal Title

      FEMS Microbilo. Lett. 243

      Pages: 393-398

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Thermostability of ancestral mutants of Caldococcus noboribetus isocitrate dehydrogenase2005

    • Author(s)
      H.Iwabata, K.Watanabe, T.Ohkuri, S.Yokobori, A.Yamagishi
    • Journal Title

      FEMS Microbilo.Lett. 243

      Pages: 393-398

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] 原始の生命体と地球の姿2004

    • Author(s)
      山岸明彦
    • Journal Title

      Biohistory 2003

      Pages: 76-79

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Life and earth in Archaean Earth (Japanese)2004

    • Author(s)
      Akihiko Yamagishi
    • Journal Title

      Biohistory 2003

      Pages: 76-79

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Evolution of Cells (Japanese)2004

    • Author(s)
      Akihiko Yamagishi
    • Journal Title

      Chemical Evolution and Cell Evolution (Iwanami)

      Pages: 9-45

    • Description
      「研究成果報告書概要(欧文)」より
  • [Book] シリーズ進化学第3巻 化学進化・細胞進化2004

    • Author(s)
      山岸明彦 他
    • Total Pages
      45
    • Publisher
      岩波書店
    • Description
      「研究成果報告書概要(和文)」より

URL: 

Published: 2007-12-13  

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