Improvement of CO2-fixing enzyme, RuBisCO, based on molecular evolution to enhance photosynthesis of plant.

2008 Fiscal Year Final Research Report

• Project/Area Number: 18688021
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Applied molecular and cellular biology
• Research Institution: Nara Institute of Science and Technology
• Principal Investigator: ASHIDA Hiroki Nara Institute of Science and Technology, バイオサイエンス研究科, 助教授 (50362851)
• Project Period (FY): 2006 – 2008
• Keywords: 植物 / 光合成 / CO2固定 / ルビスコ / 分子進化 / 紅藻

Research Abstract

ルビスコ祖先タンパク質とルビスコの比較解析から、両酵素で共通な触媒残基を明らかにした。これを基に祖先タンパク質にルビスコ触媒必須残基を導入し、試験管内でルビスコへの分子進化に成功した。紅藻ルビスコが持つ高CO_2識別残基をラン藻ルビスコに導入し、CO_2識別能を20%高めることに成功した。さらに、この残基をタバコルビスコに導入したタバコ形質転換体を作出した。紅藻ルビスコ遺伝子導入タバコの解析から、高機能外来性ルビスコの葉緑体機能発現の問題点を明らかにし、その解決策を導き出した。

Research Products

• [Journal Article] Structural and functional similarities between a RuBisCO-like protein from Bacillus subtilis and photosynthetic RuBisCO (2009)
  • Author(s): Yohtaro Saito, Hiroki Ashida, Tomoko Sakiyama, Nicole Tandeau de Marsac, Antoine Danchin, Agnieszka Sekowska, and Akiho Yokota
  • Journal Title: J. Biol. Chem. 284 Pages: 13256-13264
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary X-ray analysis of 2,3-diketo-5-methylthiopentyl-1-phosphate enolase from Bacillus subtilis (2009)
  • Author(s): Haruka Tamura, Hiroki Ashida, Shogo Koga, Yohtaro Saito, Tomonori Yadani, Yasushi Kai, Tsuyoshi Inoue, Akiho Yokota and Hiroyoshi Matsumura
  • Journal Title: Acta Crystallographica Section F65 Pages: 147-50
  • Peer Reviewed
• [Journal Article] RuBisCO-like proteins as the enolase enzyme in the methionine salvage pathway : Functional and evolutionary relationships between RuBisCO-like proteins and photosynthetic RuBisCO (2008)
  • Author(s): Hiroki Ashida, Yohtaro Saito, Toshihiro Nakano, Nicole Tandeau de Marsac, Agnieszka Sekowska, Antoine Danchin and Akiho Yokota
  • Journal Title: J. Exp. Bot. 59 Pages: 1543-54
  • Peer Reviewed
• [Journal Article] Molecular mechanisms of RuBisCO biosynthesis in higher plants (2008)
  • Author(s): Kenji Nishimura, Taro Ogawa, Hiroki Ashida, and Akiho Yokota
  • Journal Title: Plant biotech. 25 Pages: 285-290
• [Journal Article] Enzymatic characterization of 5-methylthioribulose-1-phosphate dehydratase of the methionine salvage pathway from Bacillus subtilis (2008)
  • Author(s): Hiroki Ashida, Yohtaro Saito, Chojiro Kojima and Akiho Yokota
  • Journal Title: Biosci Biotechnol Biochem. 72 Pages: 959-967
  • Peer Reviewed
• [Journal Article] Crystal structure of 5-methylthioribose 1-phosphate isomerase product complex from Bacillus subtilis : Implications for catalytic mechanism (2007)
  • Author(s): Haruka Tamura, Yohtaro Saito, Hiroki Ashida, Tsuyoshi Inoue, Yasushi Kai, Akiho Yokota, and Hiroyoshi Matsumura
  • Journal Title: Protein Sci. 17 Pages: 126-35
  • Peer Reviewed
• [Journal Article] Enzymatic Characterization of 5-Methylthioribose 1-Phosphate Isomerase from Bacillus subtilis (2007)
  • Author(s): Yohtaro Saito, Hiroki Ashida, Chojiro Kojima, Haruka Tamura, Hiroyoshi Matsumura, Yasushi Kai, Akiho Yokota
  • Journal Title: Biosci Biotechnol Biochem. 71 Pages: 2021-8
  • Peer Reviewed
• [Journal Article] A New Rubisco-like Protein Coexists with a Photosynthetic Rubisco in the Planktonic Cyanobacteria Microcystis (2006)
  • Author(s): Alyssa Carre-Mlouka, Annick Mejean, Philippe Quillardet, Hiroki Ashida, Yohtaro Saito, Akiho Yokota, Isabelle Callebaut, Agnieszka Sekowska, Elke Dittmann, Christiane Bouchier, and Nicole Tandeau de Marsac
  • Journal Title: J. Biol. Chem. 281 Pages: 24462-24471
  • Peer Reviewed
• [Presentation] Improvement of Cyanobacterial Ribulose 1,5-Bisphosphate Carboxylase/Oxygenase by Introducing the Latch Structure Involved in CO_2/O_2 Specificity in red algal RuBisCO (2008)
  • Author(s): Yohtaro Saito, Nana Ninomiya, Hiroki Ashida, and Akiho Yokota
  • Organizer: The 4^<th> INDONESIAN BIOTECHNOLOGY CONFERENCE
  • Place of Presentation: Nara Institute of Science and Technology (NAIST)
  • Year and Date: 20080805-07
• [Presentation] Analysis of common conserved residues between Bacillus RuBisCO-like protein and photosynthetic RuBisCO (2008)
  • Author(s): Toshihiro Nakano, Yohtaro Saito, Hiroki Ashida and Akiho Yokota
  • Organizer: The 4^<th> INDONESIAN BIOTECHNOLOGY CONFERENCE
  • Place of Presentation: Nara Institute of Science and Technology (NAIST)
  • Year and Date: 20080805-07
• [Presentation] RuBisCO-like proteinの研究から光合成CO_2固定酵素RuBisCOの分子進化の謎にせまる (2008)
  • Author(s): 蘆田弘樹
  • Organizer: 分子研研究会「分子の視点から見る光合成」
  • Place of Presentation: 岡崎コンファレンスセンター
  • Year and Date: 20080310-11
• [Presentation] メチオニン再生経路の酵素群によるボース新規代謝経路 (2008)
  • Author(s): 中野寿宏, 蘆田弘樹, 齋藤洋太郎, 横田明穂
  • Organizer: 日本農芸化学会2008年度大会
  • Place of Presentation: 名城大学天白キャンパス
  • Year and Date: 2008-03-27
• [Presentation] RuBisCO-like proteinの解析から見えてきたRuBisCO誕生の分子機構 (2008)
  • Author(s): 蘆田弘樹, 横田明穂
  • Organizer: 日本植物生理学会第49回年会シンポジウム 植物の生産性とカルビン回路「カルビンサイクル研究の新展開」
  • Year and Date: 2008-03-22
• [Presentation] 枯草菌RuBisCO-like proteinと光合成RuBisCOに共通する構造と機能 (2008)
  • Author(s): 齋藤洋太郎, 蘆田弘樹, Agnieszka Sekowska, Antoine Danchin, 横田明穂
  • Organizer: 第49回日本植物生理学会年会
  • Place of Presentation: 札幌
  • Year and Date: 2008-03-20
• [Presentation] メチオニン回路経路で機能する枯草菌由来RLPのX線結晶構造解析 (2007)
  • Author(s): 田村はるか, 蘆田弘樹, 斉藤洋太郎, 井上豪, 横田明穂, 甲斐泰, 松村浩由
  • Organizer: 日本結晶学会2007年度年会および総会
  • Place of Presentation: 東京
  • Year and Date: 20071201-02
• [Presentation] イネ由来RuBisCOと紅藻由来RuBisCOの硫酸イオン複合体に見られた立体構造の違い (2007)
  • Author(s): 小村泰浩, 松村浩由, 蘆田弘樹, 石田宏幸, 上野剛, 溝端栄一, 井上豪, 牧野周, 横田明穂, 前忠彦, 甲斐泰,
  • Organizer: 日本結晶学会2007年度年会および総会
  • Place of Presentation: 東京
  • Year and Date: 20071201-02
• [Presentation] Crystal structure of 5-methylthioribose 1-phosphate isomerase product complex from Bacillus subtilis : Implications for catalytic mechanism (2007)
  • Author(s): Haruka Tamura, Yohtaro Saito, Hiroki Ashida, Tsuyoshi Inoue, Yasushi Kai, Akiho Yokota, and Hiroyoshi Matsumura
  • Organizer: The 8th Conference of the Asian Crystallographic Association, AsCA' 07 第8回アジア結晶学会連合会議
  • Place of Presentation: 台北
  • Year and Date: 20071104-07
• [Presentation] STRUCTURAL DIFFERENCE BETWEEN RICE AND RED ALGA RUBISCO COMPLEXED WITH SULFATE (2007)
  • Author(s): Yasuhiro Komura, Hiroyoshi Matsumura, Hiroyuki Ishida, Hiroki Ashida, Eiichi Mizohata, Tsuyoshi Inoue, Amane Makino, Tadahiko Mae, Akiho Yokota, Yasushi Kai
  • Organizer: The 8th Conference of the Asian Crystallographic Association, AsCA' 07 第8回アジア結晶学会連合会議
  • Place of Presentation: 台北
  • Year and Date: 20071104-07
• [Presentation] Evolutionary potential of RuBisCO-like protein in Bacillus subtilis : Interaction with transition-state analogue of RuBisCO (2007)
  • Author(s): Yohtaro SAITO, Hiroki ASHIDA, Agnieszka SEKOWSKA, DANCHIN, and Akiho YOKOTA
  • Organizer: 14th International congress of photosynthesis
  • Place of Presentation: Glasgow, UK
  • Year and Date: 20070722-27
• [Presentation] Improvement of cyanobacterial RuBisCO by introducing the latch structure involved in high affinity for CO_2 in red algal RuBisCO (2007)
  • Author(s): Nana NINOMIYA, Hiroki ASHIDA, and Akiho YOKOTA
  • Organizer: 14^<th> International congress of photosynthesis
  • Place of Presentation: Glasgow, UK
  • Year and Date: 20070722-27
• [Presentation] RuBisCO-like proteins (2007)
  • Author(s): Hiroki ASHIDA and Akiho YOKOTA
  • Organizer: Research Frontiers with Rubisco - the "Elixir of Life" in the Biosphere. 14^<th> International Congress of Photosynthesis satellite Meeting
  • Place of Presentation: Harpenden, Herts, UK
  • Year and Date: 20070720-21
• [Presentation] 高CO_2親和性を示す紅藻RuBisCOが有するラッチ構造のラン藻RuBisCOへの導入 (2007)
  • Author(s): 二宮奈々, 蘆田弘樹, 横田明穂
  • Organizer: 2007年度松山年会
  • Place of Presentation: 愛媛大学城北キャンパス
  • Year and Date: 2007-03-28
• [Presentation] 枯草菌RuBisCO-like proteinsとRuBisCOの相互構造活性相関 (2007)
  • Author(s): 中野寿宏, 齋藤洋太郎, 蘆田弘樹, 横田明穂
  • Organizer: 日本農芸化学会2007年度大会
  • Place of Presentation: 東京農業大学世田谷キャンパス
  • Year and Date: 2007-03-26
• [Presentation] 光合成カルビン回路完成の分子的基盤 (2006)
  • Author(s): 蘆田弘樹, 横田明穂
  • Organizer: 日本光合成研究会第6回ワークショップセミナー New perspective of photosynthesis research 光合成研究の新たな潮流 構造とゲノム そして未来
  • Place of Presentation: 大阪大学
  • Year and Date: 20061012-13
• [Presentation] ゲノムから見たカルビン回路完成の分子機構 (2006)
  • Author(s): 蘆田弘樹, 横田明穂
  • Organizer: 日本植物学会第70回大会, シンポジウムゲノムから見た光合成機能の再発見
  • Place of Presentation: 熊本
  • Year and Date: 20060913-16
• [Presentation] X-ray structure of methylthioribose-1-phosphate isomerase from Bacillus subtilis reveals a hydride transfer mechanism of catalysis and a structure function relationship with other homologues. (2006)
  • Author(s): Haruka TAMURA, Hiroyoshi Matsumura, Yohtaro SAITO, Tsuyoshi INOUE, Hiroki ASHIDA, Akiho YOKOTA, and Yasushi KAI
  • Organizer: 20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2006-06-22
• [Presentation] Structure-function relationship between RuBisCO-like protein of Bacillus subtilis and photosynthetic RuBisCO. (2006)
  • Author(s): Yohtaro SAITO, Agnieszka SEKOWSKA, Hiroki ASHIDA, Antoine DANCHIN, and Akiho YOKOTA
  • Organizer: 20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2006-06-19
• [Presentation] Molecular evolution of RuBisCO-like proteins
  • Author(s): Hiroki Ashida, Tomoko Sakiyama, Yohtaro Saito, Nicole Tandeau de Marsac and Akiho Yokota
  • Organizer: 20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress
  • Place of Presentation: Kyoto, Japan
• [Book] Rubisco satellite meeting Research Frontiers with Rubisco, the "Elixir of Life" in the Biospherに参加して (2007)
  • Author(s): 蘆田弘樹
  • Total Pages: 84-86
  • Publisher: 光合成研究会
• [Book] プラントミメディックス〜植物に学ぶ~第2章 エネルギーと代謝 第4節 光合成CO_2固定酵素RuBisCO (2006)
  • Author(s): 蘆田弘樹, 横田明穂
  • Total Pages: 240-247
• [Patent(Industrial Property Rights)] 変異型のリブロース-1, 5-ビスリン酸カルボキシラーゼ/オキシゲナーゼおよびその利用 (2009)
  • Inventor(s): 蘆田弘樹, 横田明穂
  • Industrial Property Rights Holder: 安田國雄
  • Industrial Property Number: 特願2007-028608
  • Filing Date: 2009-02-07