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

• Project/Area Number: 19770085
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Structural biochemistry
• Research Institution: Kagawa 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: ¥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)
• Keywords: X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。

Research Products

• [Journal Article] β-D-Altrose (2009)
  • Author(s): Yuji Watanabe, Hiromi Yoshida, Kosei Takeda, Tomohiko Ishii and Shigehiro Kamitori
  • Journal Title: Acta crystallogr E 65
  • Peer Reviewed
• [Journal Article] β-D-Altrose (2009)
  • Author(s): Yuji Watanabe
  • Journal Title: Acta crystallogr E 65
  • Peer Reviewed
• [Journal Article] Overexpression, purification, crystallization and preliminary X-ray crystal analysis of Bacillus pallidus D-arabinose isomerase (2008)
  • Author(s): Kosei Takeda, Hiromi Yoshida, Goro Takada, Ken Izumori and Shigehiro Kamitori
  • Journal Title: Acta crystallogr F64(10) Pages: 589-592
  • Peer Reviewed
• [Journal Article] Overexpression, purification, crystallization and preliminary X-ray crystal analysis of Bacillus pallidus Darabinose isomerase (2008)
  • Author(s): Kosei Takeda
  • Journal Title: Acta crystallogr F 64 Pages: 589-592
  • Peer Reviewed
• [Journal Article] Crystal structures of D-tagatose 3-epimerase from Pseudomonas cichorii and its complexes with D-tagatose and D-fructose (2007)
  • 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
  • Peer Reviewed
• [Journal Article] Purification, crystallization and preliminary X-ray diffraction studies of D-tagatose 3-epimerase from Pseudomonas cichorii (2007)
  • Author(s): Hiromi Yoshida, Mitsugu Yamada, Takeyori Nishitani, Goro Takada, Ken Izumori and Shigehiro Kamitori
  • Journal Title: Acta Crystallogr F63 Pages: 123-125
  • Peer Reviewed
• [Journal Article] Crystal structures of D・tagatose 3・epimerase from Pseudomonas cichoriiand its complexes with D・tagatose and D-fructose (2007)
  • Author(s): Hiromi Yoshida
  • Journal Title: J. Mol. Biol. 374 Pages: 443-453
  • 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
• [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
• [Presentation] Crystal structure of rare sugar producing enzyme, Pseudomonas stutzeri L-rhamnose isomerase and its catalytic mechanism (2009)
  • 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
• [Presentation] Crystal structure of rare sugar producing enzyme, Pseudomonas stutzeri Lrhamnose isomerase and its catalytic mechanism (2009)
  • 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
• [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
• [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
• [Presentation] X-ray structure of D-arabinose isomerase from Bacillus pallidus (2008)
  • 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
• [Presentation] Crystal structures of rare sugars (2008)
  • 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
• [Presentation] X-ray structure of Darabinose isomerase from Bacillus pallidus (2008)
  • Author(s): Kosei Takeda (Hiromi Yoshida)
  • Organizer: Rare Sugar-Congress 2008
  • Place of Presentation: 高松
  • Year and Date: 2008-11-21
• [Presentation] Crystal structures of rare sugars (2008)
  • Author(s): Yuji Watanabe (Hiromi Ynchida)
  • Organizer: Rare Sugar Congress 2008
  • Place of Presentation: 高松
  • Year and Date: 2008-11-21
• [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 study (2008)
  • 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
• [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 study (2008)
  • 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
• [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
• [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
• [Presentation] Crystallization and preliminary X-ray analysis of D-arabinose isomerase from Bacillus pallidus (2008)
  • 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)
• [Presentation] Crystallization and preliminary X-ray analysis of Darabinose isomerase from Bacillus pallidus (2008)
  • Author(s): Kosei Takeda (Hiromi Yoshida)
  • Organizer: XXI Congress and General Assembly of the In ternational Union of Crystallography
  • Place of Presentation: 大阪
• [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
• [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
• [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
• [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
• [Presentation] Substrate-recognition mechanism of Pseudomonas stutzeri L-rhamnose isomerase based on X-ray structures and site-directed mutagenesis (2007)
  • 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
• [Presentation] SubstraterecognitionmechanismofPseudomonasstutzeriLrhamnoseisomerasebasedonXraystructuresandsitedirectedmutagenes (2007)
  • 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
• [Presentation] The Role of Metal Ion on Active Site in Pseudomonas stutzeri L-Rhamnose Isomerase (2007)
  • 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)
• [Presentation] The Role of Metal Ion on Active Site k1 Pseudomonas stutzeriL・Rhamnose Isomerase (2007)
  • Author(s): Hiromi Yoshida
  • Organizer: 32nd FEBS Congress Molecular Machines
  • Place of Presentation: ウィーン(オーストリア)
• [Remarks]
  • URL: http://www.med.kagawa-u.ac.jp/~xraylab/
• [Remarks]
  • URL: http://www.med.kagawa-u.ac.jp/xraylab/