Elucidation of novel archaeal mevalonate pathways oriented to metabolic engineering application

2013 Fiscal Year Final Research Report

• Project/Area Number: 23580111
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: Nagoya University
• Principal Investigator: HEMMI Hisashi 名古屋大学, 生命農学研究科, 准教授 (60302189)
• Project Period (FY): 2011 – 2013
• Keywords: 酵素 / 微生物 / 生体分子 / イソプレノイド / メバロン酸経路 / アーキア

Research Abstract

A part of reactions in the archaeal mevalonate pathways were unidentified. Thus we performed the conversion assay of radio-labeled substrates by cell-free extract from archaea and functional characterization of archaeal homologs of known mevalonate pathway-related enzymes. As the result, we discovered the classical mevalonate pathway, which is rare in archaea, from an archaeon of genus Sulfolobus, and a novel modified pathway, which passes through 3-phosphomevalonate, from an archaeon of genus Thermoplasma.
We also performed X-ray crystal structural studies on type 2 isopentenyl diphosphate isomerase and diphosphomevalonate decarboxylase from archaea of genus Sulfolobus. The obtained structures and the results of various analyses showed novel mechanisms of catalysis, regulation of activity, and thermostabilization.

Research Products

• [Journal Article] (R)-mevalonate 3-phosphate is an intermediate of the mevalonate pathway in Thermoplasma acidophilum (2014)
  • Author(s): Azami, Y., Hattori, A., Nishimura, H., Kawaide, H., Yoshimura, T. & Hemmi, H.
  • Journal Title: Journal of Biological Chemistry Volume: (in press)
  • Peer Reviewed
• [Journal Article] Biochemical evidence supporting the presence of the classical mevalonate pathway in the thermoacidophilic archaeon Sulfolobus solfataricus (2013)
  • Author(s): Nishimura, H., Azami, Y., Miyagawa, M., Hashimoto, C., Yoshimura, T. & Hemmi, H.
  • Journal Title: Journal of Biochemistry Volume: vol. 153 Pages: 415-420
  • Peer Reviewed
• [Journal Article] Substrate-induced change in the quaternary structure of type 2 isopentenyl diphosphate isomerase from Sulfolobus shibatae (2012)
  • Author(s): Nakatani, H., Goda, S., Unno, H., Nagai, T., Yoshimura, T. & Hemmi, H.
  • Journal Title: Journal of Bacteriology Volume: vol. 194 Pages: 3216-3224
  • Peer Reviewed
• [Journal Article] Covalent modification of reduced flavin mononucleotide in type-2 isopentenyl diphosphate isomerase by active-site-directed inhibitors (2011)
  • Author(s): Nagai, T., Unno, H., Janczak, M.W., Yoshimura, T., Poulter, C.D. & Hemmi, H.
  • Journal Title: Proceedings of the National Academy of Sciences of the United States of America Volume: vol. 108 Pages: 20461-20466
  • Peer Reviewed
• [Presentation] A third alternative mevalonate pathway from Thermoplasma acidophilum (2014)
  • Author(s): Hisashi Hemmi, Yasuhiro Azami, Ai Hattori, Hiroshi Kawaide and Tohru Yoshimura
  • Organizer: 1^<st> US-Japan Seminar on the Biosynthesis of Natural Products for Young Researchers
  • Place of Presentation: Tokyo Institute of Technology
  • Year and Date: 20140302-03
• [Presentation] 好熱性アーキア由来ジホスホメバロン酸デカルボキシラーゼの結晶構造解析 (2013)
  • Author(s): 服部愛、海野英昭、吉村徹、邊見久
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 20130911-13
• [Presentation] タイプ2イソペンテニル二リン酸イソメラーゼにおけるフラビン補酵素の触媒機能 (2013)
  • Author(s): 邊見久
  • Organizer: 2013年度日本農芸化学会大会
  • Place of Presentation: 東北大学
  • Year and Date: 20130324-27
• [Presentation] 好熱好酸性アーキアSulfolobus solfataricus由来ジホスホメバロン酸デカルボキシラーゼの立体構造 (2013)
  • Author(s): 服部愛、海野英昭、吉村徹、邊見久
  • Organizer: 第23回イソプレノイド研究会例会
  • Place of Presentation: 東京大学農学部中島董一郎ホール
  • Year and Date: 2013-09-14
• [Presentation] mechanism-based inhibitorを用いたタイプ2イソペンテニル二リン酸イソメラーゼの反応機構解析 (2012)
  • Author(s): 邊見久
  • Organizer: 2012年度酵素・補酵素研究会
  • Place of Presentation: 名古屋大学野依記念学術交流館
  • Year and Date: 20120727-28
• [Presentation] Sulfolobus solfataricusにおけるメバロン酸経路の酵素学的研究 (2012)
  • Author(s): 生明靖裕、西村泰斗、吉村徹、邊見久
  • Organizer: 日本Archaea研究会第25回講演会
  • Place of Presentation: 関西学院大学西宮上ヶ原キャンパス
  • Year and Date: 20120720-21
• [Presentation] 好熱性アーキア由来タイプ2イソペンテニル二リン酸イソメラーゼの四次構造に対する基質結合の影響 (2012)
  • Author(s): 中谷仁美、郷田秀一郎、海野英昭、長井拓也、吉村徹、邊見久
  • Organizer: 2012年度日本農芸化学会大会
  • Place of Presentation: 京都女子大学
  • Year and Date: 20120322-25
• [Presentation] 基質により誘導される好熱性アーキア由来タイプ2イソペンテニル二リン酸イソメラーゼの四次構造変化 (2012)
  • Author(s): 中谷仁美、郷田秀一郎、海野英昭、長井拓也、吉村徹、邊見久
  • Organizer: 第22回ドリコールおよびイソプレノイド研究会例会
  • Place of Presentation: 新潟大学駅南キャンパス「ときめいと」
  • Year and Date: 2012-09-29
• [Presentation] Substrate-induced change in the quaternary structure of type 2 isopentenyl diphosphate isomerase from Sulfolobus shibatae (2012)
  • Author(s): Hisashi Hemmi
  • Organizer: 新学術領域「生合成マシナリー」第4回若手シンポジウム(第8回生合成勉強会)
  • Place of Presentation: 静岡県立大学
  • Year and Date: 2012-08-25
• [Presentation] Non-redox catalysis by reduced flavin mononucleotide in type 2 isopentenyl diphosphate isomerase (2011)
  • Author(s): Hisashi Hemmi
  • Organizer: 17^<th> International Symposium on Flavins and Flavoprotein
  • Place of Presentation: University of California, Berkeley
  • Year and Date: 20110724-29
• [Presentation] 不可逆阻害剤となる基質アナログを用いたタイプ2イソペンテニル二リン酸イソメラーゼの反応機構解析 (2011)
  • Author(s): 邊見久、長井拓也、海野英昭、吉村徹
  • Organizer: 日本ビタミン学会第63回大会
  • Place of Presentation: 安田女子大学(広島市)
  • Year and Date: 20110604-05
• [Presentation] Sulfolobus属アーキアにおける古典的メバロン酸経路の酵素学的研究 (2011)
  • Author(s): 西村泰斗、橋本知佳、吉村徹、邊見久
  • Organizer: 第21回ドリコールおよびイソプレノイド研究会例会
  • Place of Presentation: 島根大学大学会館
  • Year and Date: 2011-11-04
• [Presentation] タイプ2イソペンテニル二リン酸イソメラーゼにおけるフラビン補酵素の機能 (2011)
  • Author(s): 邊見久
  • Organizer: 新学術領域「生合成マシナリー」第2回若手シンポジウム(第6回生合成勉強会)
  • Place of Presentation: 理研鈴木梅太郎ホール(和光市)
  • Year and Date: 2011-07-02
• [Book] Handbook of Flavoproteins : Volume 1–Oxidases, Dehydrogenases and Related Systems (2012)
  • Author(s): Hemmi, H. et al.
  • Total Pages: 57-73
  • Publisher: Walter de Gruyter