Cyclization mechanism analysis of terpene cyclases involved in C-C rearrangement

• Project/Area Number: 24580159
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bioproduction chemistry/Bioorganic chemistry
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: KAWAIDE Hiroshi 東京農工大学, (連合)農学研究科(研究院), 准教授 (20291916)
• 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: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 生合成 / テルペノイド / ジテルペン / 環化酵素 / NMR / 生理活性 / ditepene / Jatropha / 次世代シーケンサ

Outline of Final Research Achievements

We performed two subjects on the theses. (1) Functional analysis using isotopomers of substrate proposed a cyclization reaction mechanism of a diterpene produced by diterpene synthase from Aspergillus oryzae. Absolute configuration of the diterpene was determined by direct comparison with the same diterpene metabolite produced by another fungal metabolite. (2) Eight candidate genes encoding diterpene cyclases in Jatropha curcas were searched from our analysis data from the next generation sequencing and database published on the web. Functional analysis of the genes are in progress.
Related work was published on the Biochemical Journal.

Research Products

• [Journal Article] Molecular evolution of the substrate specificity of ent-kaurene synthases to adapt to gibberellin biosynthesis in land plants (2014)
  • Author(s): Shimane, M., Ueno, Y., Morisaki, K., Oogami, S., Natsume, M., Hayashi, K., Nozaki, H., & Kawaide, H
  • Journal Title: Biochemical Journal Volume: 462 Issue: 3 Pages: 539-546
  • DOI: 10.1042/bj20140134
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] 生合成酵素系を用いた化合物標識法による糸状菌由来新規ジテルペン環化酵素の反応解析 (2012)
  • Author(s): 菅井佳宣、嶋根真奈美、千葉康隆、三橋渉、夏目雅裕、豊増知伸、川出 洋
  • Organizer: 第54回天然有機化合物討論会
  • Place of Presentation: 東京都世田谷区（東京農業大学）
• [Presentation] 黄麹菌Aspergillus oryzae由来キメラ型新規ジテルペン合成酵素の反応解析 (2012)
  • Author(s): 菅井佳宣、嶋根真奈美、千葉康隆、三橋渉、夏目雅裕、豊増知伸、川出 洋
  • Organizer: 植物化学調節学会第47回大会
  • Place of Presentation: 山形県鶴岡市（山形大学農学部）