Germacrene A synthase in yarrow (Achillea millefolium) is an enzyme with mixed substrate specificity: gene cloning, functional characterization and expression analysis

Frontiers in Plant Science
Leila PazoukiÜlo Niinemets

Abstract

Terpenoid synthases constitute a highly diverse gene family producing a wide range of cyclic and acyclic molecules consisting of isoprene (C5) residues. Often a single terpene synthase produces a spectrum of molecules of given chain length, but some terpene synthases can use multiple substrates, producing products of different chain length. Only a few such enzymes has been characterized, but the capacity for multiple-substrate use can be more widespread than previously thought. Here we focused on germacrene A synthase (GAS) that is a key cytosolic enzyme in the sesquiterpene lactone biosynthesis pathway in the important medicinal plant Achillea millefolium (AmGAS). The full length encoding gene was heterologously expressed in Escherichia coli BL21 (DE3), functionally characterized, and its in vivo expression was analyzed. The recombinant protein catalyzed formation of germacrene A with the C15 substrate farnesyl diphosphate (FDP), while acyclic monoterpenes were formed with the C10 substrate geranyl diphosphate (GDP) and cyclic monoterpenes with the C10 substrate neryl diphosphate (NDP). Although monoterpene synthesis has been assumed to be confined exclusively to plastids, AmGAS can potentially synthesize monoterpenes in cytos...Continue Reading

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Citations

Nov 15, 2015·Applied Microbiology and Biotechnology·Mohammad MajdiMohammad Reza Abdollahi
Jul 28, 2016·Frontiers in Plant Science·Leila Pazouki, Ülo Niinemets
Jan 4, 2018·Organic & Biomolecular Chemistry·Abith VattekkatteWilhelm Boland
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Jun 16, 2021·Journal of Experimental Botany·Rachel Davidovich-RikanatiEfraim Lewinsohn
Mar 13, 2019·Organic Letters·Jan Rinkel, Jeroen S Dickschat
Sep 1, 2021·Applied Microbiology and Biotechnology·Dao-Yong GongBiao Li

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Datasets Mentioned

BETA
KC145534

Methods Mentioned

BETA
electrophoresis
PCR
transgenic

Software Mentioned

TrEMBL
MEGA
BioEdit
GenScript

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