Reaction mechanism of unique terpene synthases, that is elucidated by synthetic chemistry and structural biology
Project/Area Number |
26292058
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Research Category |
Grant-in-Aid for Scientific Research (B)
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Allocation Type | Partial Multi-year Fund |
Section | 一般 |
Research Field |
Bioorganic chemistry
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Research Institution | The University of Tokyo |
Principal Investigator |
Kuzuyama Tomohisa 東京大学, 生物生産工学研究センター, 准教授 (30280952)
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Project Period (FY) |
2014-04-01 – 2017-03-31
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Project Status |
Completed (Fiscal Year 2016)
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Budget Amount *help |
¥16,380,000 (Direct Cost: ¥12,600,000、Indirect Cost: ¥3,780,000)
Fiscal Year 2016: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2015: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2014: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
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Keywords | 生合成 / 反応機構 / テルペン / 環化酵素 / テルペノイド / 生物活性物質 / 生体触媒 / 放線菌 / 結晶構造 |
Outline of Final Research Achievements |
We identified diterpene cyclase CotB2 by sequence analysis and heterologous expression and demonstrated that CotB2 synthesizes cyclooctat-9-en-7-ol with six chiral centers from the achiral geranylgeranyl diphosphate substrate. We revealed an unusual mechanism for the exquisite CotB2-catalyzed cyclization that involves a carbon-carbon backbone rearrangement and two long-range hydrogen migrations, based on a combination of in vivo and in vitro studies with stable-isotope-labeled substrates and theoretical calculations. We also succeeded in solving the first high-resolution X-ray crystal structure of CotB2 with bound substrate analog geranylgeranyl thiodiphosphate. The combined crystal structures and mutagenesis-based biochemical assays provided structural basis for exquisite control of ring formation and stereochemistry during CotB2 catalysis. In addition, we identified and characterized a unique monoterpene synthase, cyclolavandulyl diphosphate synthase.
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Report
(4 results)
Research Products
(49 results)
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[Journal Article] "Cation-Stitching Cascade": exquisite control of terpene cyclization in cyclooctatin biosynthesis2015
Author(s)
Sato H, Teramoto K, Masumoto Y, Tezuka N, Sakai K, Ueda S, Totsuka Y, Shinada T, Nishiyama M, Wang C, Kuzuyama T, Uchiyama M
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Journal Title
Scientific Reports
Volume: 5
Pages: 18471
DOI
Related Report
Peer Reviewed / Open Access / Acknowledgement Compliant
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[Journal Article] An unusual terpene cyclization mechanism involving a carbon-carbon bond rearrangement2015
Author(s)
Meguro A, Motoyoshi Y, Teramoto K, Ueda S, Totsuka Y, Ando Y, Tomita T, Kim SY, Kimura T, Igarashi M, Sawa R, Shinada T, Nishiyama M, Kuzuyama T.
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Journal Title
Angewandte Chemie International Edition
Volume: 54
Pages: 4353-4356
DOI
Related Report
Peer Reviewed / Acknowledgement Compliant
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[Presentation] テルペンに関する最近の話題2016
Author(s)
葛山智久
Organizer
日本農芸化学会2016年度大会
Place of Presentation
札幌コンベンションセンター(北海道札幌市)
Year and Date
2016-03-27 – 2016-03-30
Related Report
Invited
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[Presentation] 放線菌由来ジテルペン環化酵素CotB2の 特異な環化反応機構の解明2015
Author(s)
寺本和矢, 目黒亜由子, 本吉祐大, 上田翔太, 遠塚悠輔, 安藤祐美, 坂井健太,富田武郎, 金承榮, 木村智之, 五十嵐雅之, 澤竜一, 品田哲郎, 西山真, 葛山智久
Organizer
イソプレノイド研究会
Place of Presentation
東北大学(宮城県仙台市)
Year and Date
2015-09-14 – 2015-09-14
Related Report
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[Presentation] アリル基質同士の縮合と環化反応を触媒する 新奇テルペン合成酵素の発見 ○小林正弥, 尾﨑太郎1, 趙平1,富田武郎, 苅田祐馬, 保野陽子, 西村栄治, 品田哲郎, 西山真, 葛山智久2015
Author(s)
小林正弥, 尾﨑太郎, 趙平, 富田武郎, 苅田祐馬, 保野陽子, 西村栄治, 品田哲郎, 西山真, 葛山智久
Organizer
イソプレノイド研究会
Place of Presentation
東北大学(宮城県仙台市)
Year and Date
2015-09-14 – 2015-09-14
Related Report
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