Revealing the mechanism of ring expansion reaction to tropolon formation uniquely found in woody plant.

• Project/Area Number: 24580248
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Wood science
• Research Institution: Kyushu University
• Principal Investigator: FUJITA Koki 九州大学, (連合)農学研究科(研究院), 助教 (90264100)
• Co-Investigator(Kenkyū-buntansha): TSUTSUMI Yuji 九州大学, 大学院農学研究院, 教授 (30236921)
• Co-Investigator(Renkei-kenkyūsha): ICHINOSE Hirofumi 九州大学, 大学院農学研究院, 准教授 (00432948)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2012: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: ヒノキチオール / トロポロン環 / テルピノレン / ＧＣＭＳ / Ｐ４５０ / ジオキシゲナーゼ / テルペン / Cupressus lusitanica / トロポロン / P450 / メキシコイトスギ / 重水素ラベル / エポキシ / 生合成 / モノテルペン

Outline of Final Research Achievements

Feeding experiment with deuterized terpinolene into Cupressus lusitanica cultured cell revealed that terpinolene is the first olefin monoterpene intermediate to biosynthesize beta-thujaplicin. This pathway was the first report that tropolon ring was formed from terpenoid.
Serching other intermediates by comparing GCMS analysis of extractives from normal and deuterized terpinolene fed to the cell did not give the good candidate of the intermediates. Therefore, we should consider another pathway driven by dioxygenase.

Research Products

• [Journal Article] Stereo-selective oxidations of terpinolene by cytochrome P450 monooxygenases in the microsomal fraction of Cupressus lusitanica cultured cells (2014)
  • Author(s): Takako Harada, Koki Fujita, Jun Shigeto, Yuji Tsutsumi
  • Journal Title: Journal of Wood Science Volume: 60 Issue: 6 Pages: 446-452
  • DOI: 10.1007/s10086-014-1432-0
  • NAID: 40020303246
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] A novel synthetic pathway for tropolone ring formation via the olefin monoterpene intermediate terpinolene in cultured Cupressus lusitanica cells (2014)
  • Author(s): Koki Fujita, Yasufumi Bunyua, Ken’ich Kurodaa, Tatsuya Ashitanib, Jun Shigetoa, Yuji Tsutsumi
  • Journal Title: Journal of Plant Physiology Volume: 171 Issue: 8 Pages: 610-614
  • DOI: 10.1016/j.jplph.2013.12.016
  • Peer Reviewed
• [Journal Article] Regio- and Substrate-specific Oxidative Metabolism of Terpinolene by Cytochrome P450 Monooxygenases in Cupressus lusitanica Cultured Cells (2012)
  • Author(s): Takako Harada, et al
  • Journal Title: American Journal of Plant Science Volume: 3-2 Issue: 02 Pages: 268-275
  • DOI: 10.4236/ajps.2012.32032
  • Peer Reviewed
• [Presentation] d6 - ラベル化テルピノレン投与による Cupressus lusitanica でのヒノキチ オール生合成中間体の探索 (2014)
  • Author(s): 藤村隼人，藤田弘毅，堤祐司
  • Organizer: 第 21 回日本木材学会 九州支部大会
  • Place of Presentation: 熊本
  • Year and Date: 2014-09-11 – 2014-09-12
• [Presentation] テルピノレンはヒノキチオール生合成中間体である (2013)
  • Author(s): 豊丹生泰史，藤田弘毅，黒田健一，芦谷竜矢，堤 祐司
  • Organizer: 第６３回日本木材学会大会（盛岡）
  • Place of Presentation: 盛岡
• [Presentation] Cuplessus lusitanica培養細胞における炭化水素モノテルペンを介したシグナル伝達 (2013)
  • Author(s): 神戸 良，藤田弘毅，堤 祐司
  • Organizer: 第６３回日本木材学会大会（盛岡）
  • Place of Presentation: 盛岡
• [Presentation] Terpinolene is the first olefin monoterpene intermediate to a tropolone, β- thujaplicin; - Potential novel pathway to tropolone ring - (2013)
  • Author(s): Koki Fujita,Yasufumi Bunyu, Ken’ichi Kuroda, Tatsuya Ashitani, Yuji Tsutsumi
  • Organizer: TERPNET 2013
  • Place of Presentation: Kolymvari, Greece
• [Presentation] The relationship between the time courses of the β-thujaplicin production and of the activities of terpinolene oxygenases in Cupressus lusitanica cultured cells (2012)
  • Author(s): TAKAKO HARADA, KOKI FUJITA AND YUJI TSUTSUMI
  • Organizer: 28th International Society of Chemical Ecology
  • Place of Presentation: Vilnius, Lithuania