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Studies on hyperthermostable aldolase : characteristics, structure and application

Research Project

Project/Area Number 15560677
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biofunction/Bioprocess
Research InstitutionThe University of Tokushima

Principal Investigator

SAKURABA Haruhiko  The University of Tokushima, Faculty of Engineering, Associate professor, 工学部, 助教授 (90205823)

Co-Investigator(Kenkyū-buntansha) OHSHIMA Toshihisa  The University of Tokushima, Faculty of Engineering, Professor, 工学部, 教授 (10093345)
Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2003: ¥1,900,000 (Direct Cost: ¥1,900,000)
KeywordsHyperthermophile / Aeropyrum pernix / Archaea / aldolase / DERA / D-2-deoxyribose-5-phosphate / aldol reaction / X-ray crystalographic analysis
Research Abstract

A gene encoding a D-2-deoxyribose-5-phosphate aldolase (DERA) homologue was identified in the hyperthermophilic archaeon Aeropyrum pernix. The gene was overexpressed in Escherichia coli, and the produced enzyme was purified and characterized. The enzyme was an extremely thermostable DERA ; the activity was not lost after incubation at 100℃ for 10 min. The enzyme had a molecular mass of about 93 kDa and consisted of four subunits with an identical molecular mass of 24 kDa. This shows the first presence of the tetrameric DERA. The three-dimensional structure of the enzyme was determined by x-ray analysis. The subunit folded into an α/β barrel. The asymmetric unit consisted of two homologous subunits, and a crystallographic 2-fold axis generated the functional tetramer. Compared with the structure of the E. coli DERA, the mainchain coordinate of the monomer of A. pernix enzyme was quite similar to that of the E. coli enzyme. A large difference in hydrophobic interactions and the number of ion pairs was not observed between the monomeric structures of the two enzymes. However, a significant difference in the quaternary structure was observed. The area of the subunit-subunit interface in the dimer of the A. pernix enzyme was much larger than the corresponding one of the E. coli enzyme. In addition, the A. pernix enzyme was 10 amino acids longer than the E. coli enzyme in the N-terminal region and exhibited an additional N-terminal helix. The N-terminal helix produced a unique dimer-dimer interface. This promotes the formation of a functional tetramer of the A. pernix enzyme and strengthens the hydrophobic inter-subunit interactions. These structural features are considered to be responsible for the extremely high stability of the A. pernix enzyme. This is the first description of the structure of hyperthermophilic DERA and of aldolase from the domain of archaea.

Report

(3 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • Research Products

    (21 results)

All 2004 2003 Other

All Journal Article (14 results) Book (1 results) Patent(Industrial Property Rights) (2 results) Publications (4 results)

  • [Journal Article] Novel archaeal Alanine : Glyoxylate aminotransferase from Thermococcus litralis.2004

    • Author(s)
      H.Sakuraba
    • Journal Title

      J Bacteriol. 186

      Pages: 5513-5518

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Unique sugar metabolism and novel enzymes of hyperthermophilic archaea.2004

    • Author(s)
      H.Sakuraba
    • Journal Title

      The Chemical Record 3

      Pages: 281-287

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Novel Archaeal Alanine ; Glyoxylate Aminotransferase from Thermococcus litoralis2004

    • Author(s)
      Haruhiko Sakuraba
    • Journal Title

      J.Bacterial. Vol.186

      Pages: 5513-5518

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Unique sugar metabolism and novel enzymes of hyperthermophilic archaea.2004

    • Author(s)
      Haruhiko Sakuraba
    • Journal Title

      The Chemical Record Vol.3

      Pages: 281-287

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Novel archaeal Alanine : Glyoxylate aminotransferase from Thermococcus litralis.2004

    • Author(s)
      H.Sakuraba
    • Journal Title

      J.Bacteriol. 186

      Pages: 5513-5518

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Unique sugar metabolism and novel enzymes of hyperthermophilic archaea.2004

    • Author(s)
      H.Sakurada
    • Journal Title

      The Chemical Record 3

      Pages: 281-287

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Cloning, Expression, and Characterization of the First Archaeal ATP-dependent Glucokinase from Aerobic Hyperthermophilic Archaeon Aeropyrum pernix2003

    • Author(s)
      H.Sakuraba
    • Journal Title

      J.Biochem. 133

      Pages: 219-224

    • NAID

      10012052941

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The First Crystal Structure of Archaeal Aldolase : Unique Tetrameric Structure of 2-De-oxy-D-ribose-5-phosphate Aldolase from Hyperthermophilic Archaea Aeropyrum pernix2003

    • Author(s)
      H.Sakuraba
    • Journal Title

      J Biol.Chem. 278

      Pages: 10799-10806

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A nicotinamide mononucleotide adenylyltransferase with unique adenylyl group donor specificity from a hyperthermophilic archaeon, Pyrococcus horikoshii OT32003

    • Author(s)
      H.Sakuraba
    • Journal Title

      J Mol.Catalys.B : Enzymatic 23

      Pages: 273-279

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The sulfolobus tokodaii gene ST1704 codes highly thermostable glucose dehydrogenase2003

    • Author(s)
      T.Ohshima
    • Journal Title

      J Mol.Catalys.B : Enzymatic 23

      Pages: 281-289

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Cloning, Expression, and Characterization of the First Archaeal ATP-dependent Glucokinase from Aerobic Hyperthermophilic Archaeon Aeropyrum pernix2003

    • Author(s)
      Haruhiko Sakuraba
    • Journal Title

      J.Biochem. Vol.133

      Pages: 219-224

    • NAID

      10012052941

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The First Crystal Structure of Archaeal Aldolase : Unique Tetrameric Structure of 2-Deoxy-D-ribose-5-phosphate Aldolase from Hyperthermophilic Archaea Aeropyrum pernix2003

    • Author(s)
      Haruhiko Sakuraba
    • Journal Title

      J.Biol.Chem. Vol.278

      Pages: 10799-10806

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A nicotinamide mononucleotide adenylyltransferase with unique adenylyl group donor specificity from a hyperthermophilic archaeon, Pyrococcus horikoshii OT32003

    • Author(s)
      Haruhiko Sakuraba
    • Journal Title

      J.Mol.Catalys.B : Enzymatic Vol.23

      Pages: 273-279

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The sulfolobus tokodaii gene ST1704 codes highly thermostable glucose dehydrogenase2003

    • Author(s)
      Toshihisa Ohshima
    • Journal Title

      J.Mol.Catalys.B : Enzymatic Vol.23

      Pages: 281-289

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] 蛋白質 核酸 酵素2003

    • Author(s)
      櫻庭 春彦
    • Total Pages
      7
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Patent(Industrial Property Rights)] キラルなヒドロキシアルデヒド化合物の製造方法2004

    • Inventor(s)
      大島 敏久 外4名
    • Industrial Property Rights Holder
      三井化学
    • Industrial Property Number
      2004-095263
    • Filing Date
      2004-03-29
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Patent(Industrial Property Rights)] キラルなヒドロキシアルデヒド化合物の製造方法2004

    • Inventor(s)
      大島敏久 外4名
    • Industrial Property Rights Holder
      三井化学
    • Industrial Property Number
      2004-095263
    • Filing Date
      2004-03-29
    • Related Report
      2004 Annual Research Report
  • [Publications] H.Sakuraba et al.: "Cloning, Expression, and Characterization of the First Archaeal ATP-dependent Glucokinase from Aerobic Hyperthermophilic Archaeon Aeropyrum pernix"J.Biochem.. 133. 219-224 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] H.Sakuraba et al.: "The First Crystal Structure of Archaeal Aldolase : Unique Tetrameric Structure of 2-Deoxy-D-ribose-5-phosphate Aldolase from Hyperthermophilic Archaea Aeropyrum pernix"J.Biol.Chem.. 278. 10799-10806 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] H.Sakuraba et al.: "A nicotinamide mononucleotide adenylyltransferase with unique adenylyl group donor specificity from a hyperthermophilic archaeon, Pyrococcus horikoshii OT3"J.Mol.Catalys.B : Enzymatic. 23. 273-279 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] T.Ohshima et al.: "The sulfolobus tokodaii gene ST1704 codes highly thermostable glucose dehydrogenase"J.Mol.Catalys.B : Enzymatic. 23. 281-289 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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