Biosynthetic enzymes-guided synthesis of intricate diterpenes

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K07407
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: 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), 教授 (20291916)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: ジテルペン / 生合成 / 植物テルペノイド / 環化酵素

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.

Free Research Field

生物有機化学