Elucidation of molecular mechanism of pyrethrin biosynthesis by a GDSL lipase and its application to creation of insect resistant plants
| Project/Area Number |
25282234
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Partial Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Biomolecular chemistry
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| Research Institution | Kinki University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
HIRATAKE Jun 京都大学, 化学研究所, 教授 (80199075)
MATSUI Kenji 山口大学, 医学(系)研究科(研究院), 教授 (90199729)
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| Co-Investigator(Renkei-kenkyūsha) |
YAMASHITA Atsuko 岡山大学, 医歯(薬)学総合研究科, 教授 (10321738)
IHARA Makoto 近畿大学, 農学部, 講師 (30466031)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥18,850,000 (Direct Cost: ¥14,500,000、Indirect Cost: ¥4,350,000)
Fiscal Year 2015: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2014: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2013: ¥10,400,000 (Direct Cost: ¥8,000,000、Indirect Cost: ¥2,400,000)
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| Keywords | 除虫菊 / ピレスリン / 生合成 / GDSLリパーゼ |
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| Outline of Final Research Achievements |
Insecticide pyrethrins containing an ester bond are biosynthesized by catalysis of a GDSL lipase TcGLIP using the acyl and the alcohol substrates. The aim of this study was to elucidate the molecular mechanism of TcGLIP in pyrethrin synthesis and regulation in terms of gene expression and catalysis. First, TcGLIP was expressed in E. coli and amino acids essential for the catalysis were identified. Next, a method to obtain recombinant TcGLIP with high yield and purity was established and solutions to crystallize the enzyme were found. On the other hand, phosphonates were synthesized as TcGLIP inhibitor candidates and tested for their inhibitory activity. As a result, several compounds were found to block the pyrethrin-synthesizing activity of TcGLIP at nanomolar concentrations. On the other hand, a TcGLIP homolog gene was knocked out in Arabidopsis, resulting in reduced potency of exudates to activate the defensin gene promoter.
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Report
(4 results)
Research Products
(22 results)
