2016 Fiscal Year Final Research Report
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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Keywords | 生合成 / 反応機構 / テルペン / 環化酵素 |
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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Free Research Field |
天然物化学
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