Campus Units

Biochemistry, Biophysics and Molecular Biology

Document Type

Article

Publication Version

Accepted Manuscript

Publication Date

9-5-2011

Journal or Book Title

ChemBioChem

Volume

12

Issue

13

First Page

1984

Last Page

1987

DOI

10.1002/cbic.201100336

Abstract

Vascular plants invariably contain a class II diterpene cyclase (EC 5.5.1.x), as an ent-copalyl diphosphate synthase is required for gibberellin phytohormone biosynthesis. This has provided the basis for evolution of a functionally diverse enzymatic family.[1] These biocatalysts fold their substrate, the general diterpenoid precursor (E,E,E)-geranylgeranyl diphosphate (GGPP), to bring the terminal three carbon-carbon double bonds into proximity with each other, and then carry out bicyclization via a protonation-initiated carbocation cascade reaction. The resulting labda-15-en-8-yl+ diphosphate intermediate is most commonly quenched by deprotonation at an exocyclic methyl, as in the production of labdadienyl/copalyl diphosphate (Scheme 1). Alternatively, the bicyclized labda-15-en-8-yl+ diphosphate intermediate can be captured by water prior to deprotonation, to form hydroxylated compounds such as labda-15-en-8-ol diphosphate.[2] In addition, this intermediate can undergo subsequent rearrangement via 1,2-hydride and/or methyl shifts, starting with the hydrogen substituent on the neighboring endocyclic methine (C9).[3] However, terminating deprotonation at the neighboring endocyclic methylene (C7) has not previously been observed. Here we report that the lycophyte Selaginella moellendorffii contains a bifunctional diterpene synthase, SmCPSKSL1, which catalyzes just such a class II cyclization reaction. In particular, SmCPSKSL1 produces an endocyclic double bond isomer of copalyl diphosphate (CPP), as well as carries out subsequent replacement of the diphosphate by a hydroxyl group to form labda-7,13E-dien-15-ol. Although this is a known plant metabolite,[4] and a small family of bioactive derived natural products is known from a phylogenetically diverse group of plants,[4-5] its biosynthesis has not been previously investigated. Our results demonstrate that this diterpenoid can be generated by a single bifunctional diterpene synthase that directly generates the endocyclic double bond, as well as hydroxyl group.

Comments

This is the peer reviewed version of the following article: Mafu, S., Hillwig, M. L. and Peters, R. J. (2011), A Novel Labda-7,13E-dien-15-ol-Producing Bifunctional Diterpene Synthase from Selaginella moellendorffii. ChemBioChem, 12: 1984–1987, which has been published in final form at doi:10.1002/cbic.201100336. This article may be used for non-commercial purposes in accordance With Wiley Terms and Conditions for self-archiving.

Copyright Owner

WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Language

en

File Format

application/pdf

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