Screening of thermophilic aldolases and application for the synthesis of sugar related materials

• Project/Area Number: 20560730
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Kagawa University
• Principal Investigator: SAKURABA Haruhiko Kagawa University, 農学部, 教授 (90205823)
• Co-Investigator(Renkei-kenkyūsha): TOSHIHISA Ohshima 九州大学, 大学院・農学研究院, 教授 (10093345)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2010: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2009: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2008: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 結晶構造解析 / 超好熱菌 / アーキア / 好熱菌 / アルドラーゼ / DERA / キメラ酵素 / PCRスワッピング / eDNA / KDGA / Sulfolobus

Research Abstract

The hyperthermophilic 2-deoxy-D-ribose-5-phosphate aldolase (DERA) gene was amplified from environmental DNA by PCR. These fragments were swapped with corresponding sequence of the known hyperthermophilic DERA gene and the expression vector for the chimera enzyme was constructed. The specific activity of the recombinant chimera DERA produced in Escherichia coli was two times higher than that of the wild type enzyme. In addition, we succeeded in determination of the crystal structure of hyperthermophilic 2-keto-3-deoxygluconate aldolases.

Research Products

• [Journal Article] Characteri-zation of a novel dye-linked L-proline dehydrogenase from anaerobic hyperthermophilic archaeon, Pyrobaculum calidifontis (2010)
  • Author(s): T.Satomura, X.Zhang, Y.Hara, K.Doi, H.Sakuraba, T.Ohshima
  • Journal Title: Appl.Microbiol.Biotech. Vol.89 Pages: 1075-1082
  • Peer Reviewed
• [Journal Article] Crystal structure of UDP-galactose 4-epimerase-like Lthreonine dehydrogenase belonging to the intermediate short-chain dehydrogenasereductase superfamily (2010)
  • Author(s): K.Yoneda, H.Sakuraba, I.Muraoka, T.Oikawa, T.Ohshima
  • Journal Title: FEBS J. Vol.277 Pages: 5124-5132
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary X-ray analysis of a novel dye-linked L-proline dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix (2010)
  • Author(s): T.Satomura, H.Sakuraba, Y.Hara, T.Ohshima
  • Journal Title: Acta.Cryst.F Vol.66 Pages: 1508-1510
  • Peer Reviewed
• [Journal Article] Catalytic properties and crystal structure of quinoprotein aldose sugar dehydrogenase from hyperthermophilic archaeon Pyrobaculum aerophilum (2010)
  • Author(s): H.Sakuraba, K.Yokono, K.Yoneda, A.Watanabe, Y.Asada, T.Satomura, T.Yabutani, J.Motonaka, T.Ohshima
  • Journal Title: Arch.Biochem.Biophys. Vol.502 Pages: 81-88
  • Peer Reviewed
• [Journal Article] The first crystal structure of L-lysine 6-dehydrogenase as an NAD-dependent amine dehydrogenase (2010)
  • Author(s): K.Yoneda, J.Fukuda, H.Sakuraba, T.Ohshima
  • Journal Title: J.Biol.Chem. Vol.285 Pages: 8444-8453
  • Peer Reviewed
• [Journal Article] Characterization of a novel dye-linked L-proline dehydrogenase from an aerobic hyperthermophilic archaeon, Pyrobaculum calidifontis (2010)
  • Author(s): Satomura, T., et al.
  • Journal Title: Appl.Microbiol.Biotech. Volume: 89 Pages: 1075-1082
  • Peer Reviewed
• [Journal Article] Crystal structure of UDP-galactose 4-epimerase-like L-threonine dehydro genase belonging to the intermediate short-chain dehydrogenase-reductase superfamily (2010)
  • Author(s): Yoneda, K., et al.
  • Journal Title: FEBS J. Volume: 277 Pages: 5124-5132
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary X-ray analysis of a novel dye-linked L-proline dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix (2010)
  • Author(s): Satomura, T., et al.
  • Journal Title: Acta.Cryst. Volume: F66 Pages: 1508-1510
  • Peer Reviewed
• [Journal Article] Catalytic properties and crystal structure of quinoprotein aldose sugar dehydrogenase from hyperthermophilic archaeon Pyrobaculum aerophilum (2010)
  • Author(s): Sakuraba, H., et al.
  • Journal Title: Arch.Biochem.Biophys. Volume: 502 Pages: 81-88
  • Peer Reviewed
• [Journal Article] The first crystal structure of L-lysine 6-dehydrogenase as an NAD-dependent amine dehydrogenase (2010)
  • Author(s): Yoneda, K., et al.
  • Journal Title: J.Biol.Chem. (in press)
  • Peer Reviewed
• [Journal Article] Structure of a D-tagatose 3-epimerase-related protein from the hyperthermophilic bacterium Thermotoga maritima (2009)
  • Author(s): H.Sakuraba, K.Yoneda, T.Satomura, R.Kawakami, T.Ohshima
  • Journal Title: Acta.Cryst.F Vol.65 Pages: 199-203
  • Peer Reviewed
• [Journal Article] Refolding, characterization and crystal structure of (S)-malatede-hydrogenase from the hyperthermophilic archaeon Aeropyrum pernix (2009)
  • Author(s): R.Kawakami, H.Sakuraba, S.Goda, H.Tsuge, T.Ohshima
  • Journal Title: Biochim.Biophys.Acta Vol.1794 Pages: 1496-1504
  • Peer Reviewed
• [Journal Article] Structure of a D-tagatose 3-epimerase-related protein from the hyperthermophilic bacterium Thermotoga maritima (2009)
  • Author(s): Sakuraba, H, et al.
  • Journal Title: Acta Crystallogr.Sect F 65 Pages: 199-203
  • Peer Reviewed
• [Journal Article] Refolding, characterization and crystal structure of (S)-malate dehydrogenase from the hyperthermophilic archaeon Aeropyrum pernix (2009)
  • Author(s): Kawakami, R., et al.
  • Journal Title: Biochim.Biophys.Acta 1794 Pages: 1496-1504
  • Peer Reviewed
• [Journal Article] 超好熱菌アルドラーゼが触媒するアルドール縮合反応 (2008)
  • Author(s): 櫻庭春彦、大島敏久
  • Journal Title: 日本生物工学会誌 Vol.86 Pages: 165-167
  • NAID: 110006668446
  • Peer Reviewed
• [Journal Article] Structure of L-aspartate oxidase from the hyperthermophilic archaeon Sulfolobus tokodaii (2008)
  • Author(s): H.Sakuraba, K.Yoneda, I.Asai, H.Tsuge, N.Katunuma, T.Ohshima
  • Journal Title: Biochim.Biophys.Acta Vol.1784 Pages: 563-571
  • Peer Reviewed
• [Journal Article] 超好熱菌アルドラーゼが触媒するアルドール縮合反応 (2008)
  • Author(s): 櫻庭春彦
  • Journal Title: 日本生物工学会誌 86 Pages: 165-167
  • NAID: 110006668446
  • Peer Reviewed
• [Journal Article] structure of L-aspartate oxidase from the hyperthermophilic archaeon Sulfolobus tokodaii. (2008)
  • Author(s): Haruhiko Sakuraba
  • Journal Title: Biochim. Biophys. Acta 1784 Pages: 563-571
  • Peer Reviewed
• [Journal Article] Crystal structure of an archaeal alanine : glyoxylate aminotransferase. (2008)
  • Author(s): Haruhiko Sakuraba
  • Journal Title: Acta Crystallographica D64 Pages: 696-699
  • Peer Reviewed
• [Presentation] 超好熱菌酵素の機能・構造解析と応用 (2010)
  • Author(s): 櫻庭春彦
  • Organizer: 日本農芸化学会中部支部第158回例会(シンポジウム)
  • Place of Presentation: 石川県女性センター
  • Year and Date: 2010-06-26
• [Presentation] 超好熱菌酵素の機能・構造解析と応用 (2010)
  • Author(s): 櫻庭春彦
  • Organizer: 日本農芸化学会中部支部第158回例会(シンポジウム)
  • Place of Presentation: 石川県女性センター(招待講演)
  • Year and Date: 2010-06-26
• [Presentation] エラープローンRCA法に基づいた耐熱性L-アスパラギン酸脱水素酵素の中性付近における触媒活性の向上 (2009)
  • Author(s): 桶崎陽友, et al.
  • Organizer: 第61回日本生物工学会大会
  • Place of Presentation: 名古屋大学
  • Year and Date: 2009-09-24
• [Presentation] 環境試料由来新規アルドラーゼ遺伝子の探索 (2008)
  • Author(s): 佐藤杏子、櫻庭春彦、大島敏久
  • Organizer: 日本農芸化学会中四国支部2008年度支部大会
  • Place of Presentation: 鳥取大学・農学部
  • Year and Date: 2008-09-13
• [Presentation] 環境試料由来新規アルドラーゼ遺伝子の探索 (2008)
  • Author(s): 佐藤杏子
  • Organizer: 日本農芸化学会中四国支部2008年度文部大会
  • Place of Presentation: 鳥取大学・農学部
  • Year and Date: 2008-09-13
• [Remarks] ホームページ等
  • URL: http://www.ag.kagawa-u.ac.jp/sakuraba/index.html
• [Remarks]
  • URL: http://www.ag.kagawa-u.ac.jp/sakuraba/index.html