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Crystal structure analysis of rare sugar producible enzymes for their effective production by protein engineering

Research Project

Project/Area Number 19770085
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Structural biochemistry
Research InstitutionKagawa University

Principal Investigator

YOSHIDA Hiromi  Kagawa University, 総合生命科学研究センター, 准教授 (10313305)

Project Period (FY) 2007 – 2008
Project Status Completed (Fiscal Year 2008)
Budget Amount *help
¥3,660,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥360,000)
Fiscal Year 2008: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2007: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsX線結晶解析 / 糖異性化酵素 / L-ラムノースイソメラーゼ / D-タガトースエピメラーゼ / D-アラビノースイソメラーゼ / 希少糖 / 放射光 / 部位特異的変異
Research Abstract

天然には微量にしか存在しない「希少糖」は糖としての食品分野への応用だけではなく医療分野においても希少糖の利用価値が期待されている。本研究は「希少糖」の生産に利用される酵素のより効率的な生産を目指した分子設計に有用な情報を得ることを目的に、希少糖生産酵素の構造・機能解析を行った。本研究において、Pseudomonas stutzeri由来L-ラムノースイソメラーゼについては変異酵素を触媒機構に関する詳細な情報が得られ、Pseudomonas cichorii由来D-タガトース3-エピメラーゼとBacillus pallidus由来D-アラビノースイソメラーゼの構造を新たに決定した。

Report

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

    (33 results)

All 2009 2008 2007 Other

All Journal Article (7 results) (of which Peer Reviewed: 7 results) Presentation (24 results) Remarks (2 results)

  • [Journal Article] β-D-Altrose2009

    • Author(s)
      Yuji Watanabe, Hiromi Yoshida, Kosei Takeda, Tomohiko Ishii and Shigehiro Kamitori
    • Journal Title

      Acta crystallogr E 65

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] β-D-Altrose2009

    • Author(s)
      Yuji Watanabe
    • Journal Title

      Acta crystallogr E 65

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Overexpression, purification, crystallization and preliminary X-ray crystal analysis of Bacillus pallidus D-arabinose isomerase2008

    • Author(s)
      Kosei Takeda, Hiromi Yoshida, Goro Takada, Ken Izumori and Shigehiro Kamitori
    • Journal Title

      Acta crystallogr F64(10)

      Pages: 589-592

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Overexpression, purification, crystallization and preliminary X-ray crystal analysis of Bacillus pallidus Darabinose isomerase2008

    • Author(s)
      Kosei Takeda
    • Journal Title

      Acta crystallogr F 64

      Pages: 589-592

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystal structures of D-tagatose 3-epimerase from Pseudomonas cichorii and its complexes with D-tagatose and D-fructose2007

    • Author(s)
      Hiromi Yoshida, Mitsugu Yamada, Takeyori Nishitani, Goro Takada, Ken Izumori and Shigehiro Kamitori
    • Journal Title

      J. Mol. Biol 374(2)

      Pages: 443-453

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Purification, crystallization and preliminary X-ray diffraction studies of D-tagatose 3-epimerase from Pseudomonas cichorii2007

    • Author(s)
      Hiromi Yoshida, Mitsugu Yamada, Takeyori Nishitani, Goro Takada, Ken Izumori and Shigehiro Kamitori
    • Journal Title

      Acta Crystallogr F63

      Pages: 123-125

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Crystal structures of D・tagatose 3・epimerase from Pseudomonas cichoriiand its complexes with D・tagatose and D-fructose2007

    • Author(s)
      Hiromi Yoshida
    • Journal Title

      J. Mol. Biol. 374

      Pages: 443-453

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Presentation] 変異酵素の結晶解析に基づくPseudomonas stutzeri由来L-ラムノースイソメラーゼにおけるC末端領域の役割2009

    • Author(s)
      吉田裕美,武田耕青,何森健,神鳥成弘
    • Organizer
      2009年度農芸化学会大会
    • Place of Presentation
      (福岡)
    • Year and Date
      2009-03-29
    • Related Report
      2008 Final Research Report
  • [Presentation] 変異酵素の結晶解析に基づくPseudomonas stutzeri由来L-ラムノースイソメラーゼにおけるC末端領域の役割2009

    • Author(s)
      吉田裕美
    • Organizer
      2009年度農芸化学会大会
    • Place of Presentation
      福岡
    • Year and Date
      2009-03-29
    • Related Report
      2008 Annual Research Report
  • [Presentation] Crystal structure of rare sugar producing enzyme, Pseudomonas stutzeri L-rhamnose isomerase and its catalytic mechanism2009

    • Author(s)
      Hiromi Yoshida, Mitsugu Yamada, Goro Takada, Ken Izumori and Shigehiro Kamitori
    • Organizer
      38th National Seminar on Crystallography
    • Place of Presentation
      (Mysore, India)
    • Year and Date
      2009-02-13
    • Related Report
      2008 Final Research Report
  • [Presentation] Crystal structure of rare sugar producing enzyme, Pseudomonas stutzeri Lrhamnose isomerase and its catalytic mechanism2009

    • Author(s)
      Hiromi Yoshida
    • Organizer
      38th National Seminar on Crystallography
    • Place of Presentation
      マイソール(インド)
    • Year and Date
      2009-02-13
    • Related Report
      2008 Annual Research Report
  • [Presentation] Pseudomonas stutzeri L-ラムノースイソメラーゼの触媒反応機構に関与するアミノ酸残基改変型酵素のX線結晶構造解析2008

    • Author(s)
      吉田裕美,渡部優史,武田耕青,石井知彦,何森健,神鳥成弘
    • Organizer
      第31回日本分子生物学会年会第81回日本生化学会大会合同大会
    • Place of Presentation
      (神戸)
    • Year and Date
      2008-12-11
    • Related Report
      2008 Final Research Report
  • [Presentation] Pesudomonas stutzeri L-ラムノースイソメラーゼの触媒反応機構に関与するアミン酸残基改変型酵素のX線結晶構造解析2008

    • Author(s)
      吉田裕美
    • Organizer
      第31回日本分子生物学会年会第81回日本生化学会大会合同大会
    • Place of Presentation
      神戸
    • Year and Date
      2008-12-11
    • Related Report
      2008 Annual Research Report
  • [Presentation] X-ray structure of D-arabinose isomerase from Bacillus pallidus2008

    • Author(s)
      Kosei Takeda, Hiromi Yoshida, Kenji Morimoto, Goro Takada, Ken Izumori and Shigehiro Kamitori
    • Organizer
      Rare Sugar Congress 2008
    • Place of Presentation
      (Takamatsu, Japan)
    • Year and Date
      2008-11-21
    • Related Report
      2008 Final Research Report
  • [Presentation] Crystal structures of rare sugars2008

    • Author(s)
      Yuji Watanabe, Kosei Takeda, Hiromi Yoshida, Tomohiko Ishii, Goro Takada, Ken Izumori and Shigehiro Kamitori
    • Organizer
      Rare Sugar Congress 2008
    • Place of Presentation
      (Takamatsu, Japan)
    • Year and Date
      2008-11-21
    • Related Report
      2008 Final Research Report
  • [Presentation] X-ray structure of Darabinose isomerase from Bacillus pallidus2008

    • Author(s)
      Kosei Takeda (Hiromi Yoshida)
    • Organizer
      Rare Sugar-Congress 2008
    • Place of Presentation
      高松
    • Year and Date
      2008-11-21
    • Related Report
      2008 Annual Research Report
  • [Presentation] Crystal structures of rare sugars2008

    • Author(s)
      Yuji Watanabe (Hiromi Ynchida)
    • Organizer
      Rare Sugar Congress 2008
    • Place of Presentation
      高松
    • Year and Date
      2008-11-21
    • Related Report
      2008 Annual Research Report
  • [Presentation] The role of Cys66 of D-tagatose 3-epimerase from Pseudomonas cichorii on the basis of the crystal structure and its site-directed mutagenesis study2008

    • Author(s)
      Hiromi Yoshida, Kosei Takeda, Goro Takada, Ken Izumori and Shigehiro Kamitori
    • Organizer
      XXIV International Carbohydrate Symposium
    • Place of Presentation
      (Oslo, Norway)
    • Year and Date
      2008-07-29
    • Related Report
      2008 Final Research Report
  • [Presentation] The role of Cys66 of D-tagatose 3-epimerase from Pseudomonas cichorii on the basis of the crystal structure and its site-directed mutagenesis study2008

    • Author(s)
      Hiromi Yoshida
    • Organizer
      XXIV International Carbohydrate Symposium
    • Place of Presentation
      オイロ(ノルウェー)
    • Year and Date
      2008-07-29
    • Related Report
      2008 Annual Research Report
  • [Presentation] X線結晶解析に基づくPseudomonas stutzeri由来L-ラムノースイソメラーゼにおけるSer329の基質認識への影響2008

    • Author(s)
      吉田裕美,武田耕青,高田悟郎,何森健,神鳥成弘
    • Organizer
      2008年度農芸化学会大会
    • Place of Presentation
      (名古屋)
    • Year and Date
      2008-03-28
    • Related Report
      2008 Final Research Report
  • [Presentation] X線結晶解析に基づくPseudomonas stutzeri由来L・ラムノースイソメラーゼにおけるSer329の基質認識への影響2008

    • Author(s)
      吉田裕美
    • Organizer
      2008年度農芸化学会大会
    • Place of Presentation
      名古屋
    • Year and Date
      2008-03-28
    • Related Report
      2007 Annual Research Report
  • [Presentation] Crystallization and preliminary X-ray analysis of D-arabinose isomerase from Bacillus pallidus2008

    • Author(s)
      Kosei Takeda, Hiromi Yoshida, Goro Takada, Ken Izumori and Shigehiro Kamitori
    • Organizer
      XXI Congress and General Assembly of the International Union of Crystallography
    • Place of Presentation
      (Osaka, Japan)
    • Related Report
      2008 Final Research Report
  • [Presentation] Crystallization and preliminary X-ray analysis of Darabinose isomerase from Bacillus pallidus2008

    • Author(s)
      Kosei Takeda (Hiromi Yoshida)
    • Organizer
      XXI Congress and General Assembly of the In ternational Union of Crystallography
    • Place of Presentation
      大阪
    • Related Report
      2008 Annual Research Report
  • [Presentation] seudomonas stutzeriL-ラムノースイソメラーゼ変異酵素S329Fの構造解析と基質結合様式2007

    • Author(s)
      吉田裕美,武田耕青,高田悟郎,何森健,神鳥成弘
    • Organizer
      第30回日本分子生物学会年会第80回日本生化学会大会合同大会
    • Place of Presentation
      (横浜)
    • Year and Date
      2007-12-11
    • Related Report
      2008 Final Research Report
  • [Presentation] Pseudomonas stutzeri L・ラムノースイソメラーゼ変異酵素S329Fの構造解析と基質結合様式2007

    • Author(s)
      吉田裕美
    • Organizer
      第30回日本分子生物学会年会第80回日本生化学会大会合同大会
    • Place of Presentation
      横浜
    • Year and Date
      2007-12-11
    • Related Report
      2007 Annual Research Report
  • [Presentation] 土壌菌D-タガトースエピメラーゼの構造と触媒反応機構2007

    • Author(s)
      吉田裕美,武田耕青,高田悟郎,何森健,神鳥成弘
    • Organizer
      2007年度酵素・補酵素研究会
    • Place of Presentation
      (倉敷)
    • Year and Date
      2007-11-09
    • Related Report
      2008 Final Research Report
  • [Presentation] 土壌菌D・タガトースェピメラーゼの構造と触媒反応機構2007

    • Author(s)
      吉田裕美
    • Organizer
      2007年度酵素・補酵素研究会
    • Place of Presentation
      横浜
    • Year and Date
      2007-11-09
    • Related Report
      2007 Annual Research Report
  • [Presentation] Substrate-recognition mechanism of Pseudomonas stutzeri L-rhamnose isomerase based on X-ray structures and site-directed mutagenesis2007

    • Author(s)
      Kosei Takeda, Hiromi Yoshida, Goro Takada, Ken Izumori Tomohiko Ishii and Shigehiro Kamitori
    • Organizer
      2nd International Symposium on Diffraction Structural Biology
    • Place of Presentation
      (Tokyo, Japan)
    • Year and Date
      2007-09-11
    • Related Report
      2008 Final Research Report
  • [Presentation] SubstraterecognitionmechanismofPseudomonasstutzeriLrhamnoseisomerasebasedonXraystructuresandsitedirectedmutagenes2007

    • Author(s)
      Kosei Takeda (Hiromi Yoshida)
    • Organizer
      2nd International Symposium on Diffraction Structural Biology
    • Place of Presentation
      東京
    • Year and Date
      2007-09-11
    • Related Report
      2007 Annual Research Report
  • [Presentation] The Role of Metal Ion on Active Site in Pseudomonas stutzeri L-Rhamnose Isomerase2007

    • Author(s)
      Hiromi Yoshida, Masatsugu Yamaji, Mitsugu Yamada, Goro Takada, Ken Izumori Tomohiko Ishii and Shigehiro Kamitori
    • Organizer
      32nd FEBS Congress Molecular Machine
    • Place of Presentation
      (Vienna, Austria)
    • Related Report
      2008 Final Research Report
  • [Presentation] The Role of Metal Ion on Active Site k1 Pseudomonas stutzeriL・Rhamnose Isomerase2007

    • Author(s)
      Hiromi Yoshida
    • Organizer
      32nd FEBS Congress Molecular Machines
    • Place of Presentation
      ウィーン(オーストリア)
    • Related Report
      2007 Annual Research Report
  • [Remarks]

    • URL

      http://www.med.kagawa-u.ac.jp/~xraylab/

    • Related Report
      2008 Final Research Report
  • [Remarks]

    • URL

      http://www.med.kagawa-u.ac.jp/xraylab/

    • Related Report
      2008 Annual Research Report

URL: 

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

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