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Biosynthetic enzymes-guided synthesis of intricate diterpenes

Research Project

Project/Area Number 15K07407
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Bioorganic chemistry
Research InstitutionTokyo 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), 教授 (20291916)

Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywordsジテルペン / 生合成 / 植物テルペノイド / 環化酵素 / Euphorbiaceae / diterpene / biosynthesis / cyclase / ユーフォルビア属 / ジテルペン環化酵素 / Euphorbia
Outline of Final Research Achievements

Euphorbiaceae plants produce potent bioactive diterpenoid metabolites such as phorbol esters and ingenol-related compounds. To determine biosynthetic enzyme genes involved in phorbol and ingenol biosyntheses, two species (Euphorbia milli and Euphorbia tirucalli) was used as plant materials for analysis of mRNA expression by next-generation sequencer (RNAseq analysis). RNAseq analyses for screening of mRNAs encoding terpene-biosynthetic enzymes detected eleven sequences and five sequences from E. milli and E. turicalli, respectively. Based on the analyses of BLAST and HMMER searches, we isolated four genes (EmDTC-15, EmDTC-16, EmDTC-23 and EmDTC-24) and three genes (EtDTC-1, EtDTC-4 and EtDTC-5) as putative diterpene cyclase genes from E. milli and E. turicalli, respectively. As the results of functional analysis, one gene, EtDTC-1, encodes ent-kaurene synthase, and another gene, EmDTC-24 encodes an unknown diterpene cyclase that recognizes ent-copalyl diphosphate as a substrate.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report

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Published: 2015-04-16   Modified: 2019-03-29  

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