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Promotion of reverse reaction of carbohydrolase and its applicationfor oligosaccharide synthesis

Research Project

Project/Area Number 19580102
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied biochemistry
Research InstitutionHokkaido University

Principal Investigator

MORI Aruhide  Hokkaido University, 大学院・農学研究院, 准教授 (80241363)

Project Period (FY) 2007 – 2008
Project Status Completed (Fiscal Year 2008)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2008: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2007: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Keywords酵素利用学 / オリゴ糖 / 加水分解酵素 / 逆反応 / オリゴ糖合成 / アノマー反転型酵素
Research Abstract

トレハラーゼはトレハロースを加水分解する.本研究では, トレハラーゼ改変酵素と特殊化合物(βフッ化グルコース)を用いて, 加水分解の逆反応によりトレハロースを高効率で合成させることに成功した.変異酵素として, 塩基触媒変異体, および塩基触媒および加水分解の基質の水分子に影響を与えるアミノ酸残基変異体を用いた.何れも反応速度は野生型に比べ低下したが, 特に, 後者の合成効率が高く, 60%程度の収率を示した.

Report

(3 results)
  • 2008 Annual Research Report   Final Research Report ( PDF )
  • 2007 Annual Research Report
  • Research Products

    (26 results)

All 2009 2008 2007

All Journal Article (11 results) (of which Peer Reviewed: 11 results) Presentation (14 results) Book (1 results)

  • [Journal Article] Structural and Functional Analysis of aGlycoside Hydrolase Family 97 Enzyme fromBacteroides thetaiotaomicron2008

    • Author(s)
      Kitamura M, Okuyama M
    • Journal Title

      J. Biol. Chem. 283(52)

      Pages: 6328-6337

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Structure-function relationship of substrate lengthspecificity of dextran glucosidase from Streptococcusmutants Biologia2008

    • Author(s)
      Saburi W, Hondoh H
    • Journal Title

      63(6)

      Pages: 1000-1005

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Substrate recognition mechanism ofα-1,6-glucosidic linkage hydrolyzing enzyme,dextran glucosidase from Streptococcus mutans2008

    • Author(s)
      Hondoh H, Saburi W, Mori H
    • Journal Title

      J Mol Biol. 378(4)

      Pages: 911-920

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Structural and Functional Analysis of a Glycoside Hydrolase Family 97 Enzyme from Bacteroides thetaiotaomicron2008

    • Author(s)
      Kitamura M
    • Journal Title

      J. Biol. Chem 283

      Pages: 6328-6337

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A. Substrate recognitlon mechanism of α-1,6-glucosi4ic linkage hydrolyzing enzyme, dextran giucosidase from Streptococcus mutans2008

    • Author(s)
      Hondoh H
    • Journal Title

      J. Mol. Biol 378

      Pages: 911-920

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Aglycone specificity of Escherichia coliα-xylosidase investigated by transxylosylation2007

    • Author(s)
      Kang MS, Okuyama M
    • Journal Title

      FEBS J. 274(23)

      Pages: 6074-6084

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Function-unknown glycoside hydrolase family31 proteins, mRNAs of which were expressed inrice ripening and germinating stages, areα-glucosidase and α-xylosidase. J Biochem2007

    • Author(s)
      Nakai H, Tanizawa S,
    • Journal Title

      J Biochem. 142(4)

      Pages: 491-500

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Crystallization and preliminary X-ray analysis ofStreptococcus mutans dextran glucosidase2007

    • Author(s)
      Saburi W, Hondoh H
    • Journal Title

      Acta Crystallogr Sect F Struct Biol CrystCommun 63(Pt9)

      Pages: 774-776

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Molecular Cloning of cDNA for Trehalase fromthe European Honeybee, Apis mellifera L., andIts Heterologous Expression in Pichia pastoris2007

    • Author(s)
      Lee, J.-H., Saito, S., Mori, H
    • Journal Title

      Biosci. Biotechnol. Biochem 71(9)

      Pages: 2256-2265

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Molecular Cloning of cDNAs and Genes forThree α-Glucosidases from European Honeybees,Apis mellifera L., and Heterologous Productionof Recombinant Enzymes in Pichia pastoris2007

    • Author(s)
      Nishimoto M, Mori H
    • Journal Title

      Biosci. Biotechnol. Biochem 71(7)

      Pages: 1703-1716

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Cloning of cDNA for Trehalase from the European Honeybee, Apis mellifera L., and Its Heterologous Expressiong in Pichia pastoris.2007

    • Author(s)
      Lee, J. -H.
    • Journal Title

      Biosci. Biotechnol. Biochem. 71

      Pages: 2256-2265

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Presentation] 加水分解反応の抑制によりトレハロースを縮合蓄積させるトレハラーゼ変異酵素2009

    • Author(s)
      牧 孝多朗
    • Organizer
      日本農芸化学会大会
    • Place of Presentation
      福岡
    • Year and Date
      2009-03-28
    • Related Report
      2008 Final Research Report
  • [Presentation] Isomaltooligosaccharide 6-α-glucosyltransferase(I6GT)のイソマルトオリゴ糖への作用と転移に関する構造因子2009

    • Author(s)
      西村崇志
    • Organizer
      日本農芸化学会大会
    • Place of Presentation
      福岡
    • Year and Date
      2009-03-28
    • Related Report
      2008 Final Research Report
  • [Presentation] 加水分解反応の抑制によりトレハロースを縮合蓄積させるトレハラーゼ変異酵素2009

    • Author(s)
      牧孝太朗
    • Organizer
      日本農芸化学会大会
    • Place of Presentation
      マリンメッセ(福岡)
    • Year and Date
      2009-03-28
    • Related Report
      2008 Annual Research Report
  • [Presentation] 大腸菌trehalase の触媒アミノ酸残基の決定とglycosynthase 反応2008

    • Author(s)
      森春英
    • Organizer
      日本応用糖質科学会大会
    • Place of Presentation
      沖縄
    • Year and Date
      2008-09-18
    • Related Report
      2008 Final Research Report
  • [Presentation] デキストリンデキストラナーゼ遺伝子の異種宿主発現および組換え酵素の解析2008

    • Author(s)
      貞廣樹里
    • Place of Presentation
      沖縄
    • Year and Date
      2008-09-18
    • Related Report
      2008 Final Research Report
  • [Presentation] 大腸菌trehalaseの触媒アミノ酸残基の決定とglycosynthase反応2008

    • Author(s)
      森春英
    • Organizer
      日本応用糖質科学会大会
    • Place of Presentation
      琉球大(那覇・沖縄)
    • Year and Date
      2008-09-18
    • Related Report
      2008 Annual Research Report
  • [Presentation] 新規酵素isomaltooligosaccharide 6-α-glucosyltransferase(I6GT)の機能解析2008

    • Author(s)
      鐘ヶ江倫世
    • Organizer
      日本農芸化学会北海道支部学術合同講演会
    • Place of Presentation
      札幌
    • Year and Date
      2008-08-08
    • Related Report
      2008 Final Research Report
  • [Presentation] デキストリンデキストラナーゼの一次構造決定および組換え酵素の解析2008

    • Author(s)
      貞廣樹里
    • Organizer
      日本農芸化学会北海道支部講演会
    • Place of Presentation
      北海道大学(札幌)
    • Year and Date
      2008-08-08
    • Related Report
      2008 Annual Research Report
  • [Presentation] Identification of essential catalyticresidues in an Escherichia coli trehalase and aninverting glycosynthase reaction catalyzed by thecatalytic base-mutated enzyme2008

    • Author(s)
      Mori H
    • Organizer
      XXIVInternational Carbohydrate Symposium
    • Place of Presentation
      オスロ大学
    • Year and Date
      2008-07-28
    • Related Report
      2008 Final Research Report
  • [Presentation] Identification of essential catalytic residues in an Escherichia coli trehalase and an inverting glycosynthase reaction catalyzed by the catalytic base-mutated enzyme2008

    • Author(s)
      森春英
    • Organizer
      International Carbohydrate Symposium(国際炭水化物シンポジウム)
    • Place of Presentation
      ホスロ大学(オスロ,ノルウェー)
    • Year and Date
      2008-07-28
    • Related Report
      2008 Annual Research Report
  • [Presentation] Functional and structural analysis of glycoside hydrolase family 97 enzyme from Bacteroides thetaiotaomicron2008

    • Author(s)
      奥山正幸
    • Organizer
      International Carbohydrate Symposium(国際炭水化物シンポジウム)
    • Place of Presentation
      オスロ大学(オスロ,ノルウェー)
    • Year and Date
      2008-07-28
    • Related Report
      2008 Annual Research Report
  • [Presentation] 新しく見出されたイソマルトオリゴ糖不均化酵素の遺伝子クローニングと異種宿主発現2008

    • Author(s)
      鐘ケ江 倫世
    • Organizer
      日本農芸化学会大会
    • Place of Presentation
      名古屋(名城大)
    • Year and Date
      2008-03-28
    • Related Report
      2008 Final Research Report
  • [Presentation] Functional and structural analysisof glycoside hydrolase family 97 enzyme fromBacteroides thetaiotaomicron2008

    • Author(s)
      Okuyama M
    • Organizer
      XXIV InternationalCarbohydrate Symposium
    • Place of Presentation
      オスロ大学
    • Related Report
      2008 Final Research Report
  • [Presentation] Substrate recognition mechanism ofdextran glucosidase from Streptococcus mutans(DexB)2007

    • Author(s)
      Hondoh H
    • Organizer
      Carbohydrate Bioengineering Meeting 7
    • Place of Presentation
      ドイツ/ブラウンシュヴァイ
    • Related Report
      2008 Final Research Report
  • [Book] Molecular mechanism ofα-glucosidase In Carbohydrate-active enzymes:structure, function, and applications2008

    • Author(s)
      Okuyama M, Mori H, Hondoh H, Nakai H, SaburiW, Kang MS, Kim YM, Nishimoto M, WongchawalitJ, Yamamoto T, Son M, Lee JH, Mar SS, Fukuda K,Chiba S, Kimura A
    • Publisher
      Woodhead Publishing Ltd.,Cambridge, UK
    • Related Report
      2008 Final Research Report

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Published: 2007-04-01   Modified: 2016-04-21  

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