Enzymatic 13C Labeling and Multidimensional NMR Analysis of Miltiradiene Synthesized by Bifunctional Diterpene Cyclase in Selaginella moellendorffii

Sugai, Yoshinori; Ueno, Yoshimitsu; Hayashi, K.; Oogami, Shingo; Toyomasu, Tomonobu; Matsumoto, Sadamu; Natsume, Masahiro; Nozaki, Hirōshi; Kawaide, Hiroshi

doi:10.1074/jbc.m111.302703

Cited by 42 publications

(43 citation statements)

References 32 publications

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“…As monofunctional diterpene synthases, they convert GGPP to copalyl diphosphate, which is the substrate for gibberellins or other diterpenoids. In contrast, the previously characterized SmTPS7 and SmTPS4 function as bifunctional diterpene synthases, catalyzing the consecutive reactions of GGPP to copalyl diphosphate to final terpene products (20,21). These results indicate that S. moellendorffii contains both bifunctional and monofunctional diterpene synthases.…”

Section: Discussioncontrasting

confidence: 48%

Nonseed plant Selaginella moellendorffii has both seed plant and microbial types of terpene synthases

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Yin³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

110

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Terpene synthases (TPSs) are pivotal enzymes for the biosynthesis of terpenoids, the largest class of secondary metabolites made by plants and other organisms. To understand the basis of the vast diversification of these enzymes in plants, we investigated Selaginella moellendorffii , a nonseed vascular plant. The genome of this species was found to contain two distinct types of TPS genes. The first type of genes, which was designated as S. moellendorffii TPS genes ( SmTPSs ), consists of 18 members. SmTPSs share common ancestry with typical seed plant TPSs . Selected members of the SmTPSs were shown to encode diterpene synthases. The second type of genes, designated as S. moellendorffii microbial TPS -like genes ( SmMTPSLs ), consists of 48 members. Phylogenetic analysis showed that SmMTPSLs are more closely related to microbial TPSs than other plant TPSs. Selected SmMTPSLs were determined to function as monoterpene and sesquiterpene synthases. Most of the products formed were typical monoterpenes and sesquiterpenes that have been previously shown to be synthesized by classical plant TPS enzymes. Some in vitro products of the characterized SmMTPSLs were detected in the headspace of S. moellendorffii plants treated with the fungal elicitor alamethicin, showing that they are also formed in the intact plant. The presence of two distinct types of TPSs in the genome of S. moellendorffii raises the possibility that the TPSs in other plant species may also have more than one evolutionary origin.

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Section: Discussioncontrasting

confidence: 48%

Nonseed plant Selaginella moellendorffii has both seed plant and microbial types of terpene synthases

Köllner

Yin³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

110

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“…Bifunctional CPS-KSs have been found in nonvascular plants, although that from P. patens, a non-seed ancestral moss, produces largely 16a-hydroxy-ent-kaurane (Hayashi et al, 2006), while that from the liverwort J. subulata produces exclusively ent-kaurene . S. moellendorffii, a lycophyte occupying an intermediate position in the evolutionary landscape (Figure 1) between bryophytes and the euphyllophytes also has been reported to possess bifunctional diterpene synthases, although neither of those characterized to date produces ent-kaurene (Mafu et al, 2011;Sugai et al, 2011). S. moellendorffii also contains monofunctional CPSs (Li et al, 2012), one of which produces ent-CPP, as well as a monofunctional KS (Shimane et al, 2014).…”

Section: Diterpene Synthases Provide An Anchor For Assessmentmentioning

confidence: 99%

Molecular Diversity of Terpene Synthases in the Liverwort Marchantia polymorpha

et al. 2016

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Marchantia polymorpha is a basal terrestrial land plant, which like most liverworts accumulates structurally diverse terpenes believed to serve in deterring disease and herbivory. Previous studies have suggested that the mevalonate and methylerythritol phosphate pathways, present in evolutionarily diverged plants, are also operative in liverworts. However, the genes and enzymes responsible for the chemical diversity of terpenes have yet to be described. In this study, we resorted to a HMMER search tool to identify 17 putative terpene synthase genes from M. polymorpha transcriptomes. Functional characterization identified four diterpene synthase genes phylogenetically related to those found in diverged plants and nine rather unusual monoterpene and sesquiterpene synthase-like genes. The presence of separate monofunctional diterpene synthases for ent-copalyl diphosphate and ent-kaurene biosynthesis is similar to orthologs found in vascular plants, pushing the date of the underlying gene duplication and neofunctionalization of the ancestral diterpene synthase gene family to >400 million years ago. By contrast, the mono-and sesquiterpene synthases represent a distinct class of enzymes, not related to previously described plant terpene synthases and only distantly so to microbial-type terpene synthases. The absence of a Mg 2+ binding, aspartate-rich, DDXXD motif places these enzymes in a noncanonical family of terpene synthases.

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“…Class I diTPSs, which harbor DDxxD and NSE/DTE functional motifs in the a-domain, then catalyze the ionization of the diphosphate ester and subsequent rearrangement reactions (Xu et al, 2007a;Gao et al, 2009;Keeling et al, 2010;Caniard et al, 2012;Sallaud et al, 2012). In contrast, the archetypical diTPSs of nonvascular plants, such as Physcomitrella patens or Jungermannia subulata (Hayashi et al, 2006;Kawaide et al, 2011), the specialized diTPSs of the lycophyte Selaginella moellendorffii (Mafu et al, 2011;Sugai et al, 2011), and the gymnosperm diTPSs involved in diterpene resin acid (DRA) metabolism (Vogel et al, 1996;Peters et al, 2000;Schepmann et al, 2001;Martin et al, 2004;Ro and Bohlmann, 2006;Keeling et al, 2011a, Figure 1. diTPS-catalyzed biosynthesis of general and specialized diterpenes in conifers.…”

mentioning

confidence: 99%

Evolution of Conifer Diterpene Synthases: Diterpene Resin Acid Biosynthesis in Lodgepole Pine and Jack Pine Involves Monofunctional and Bifunctional Diterpene Synthases

et al. 2012

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Diterpene resin acids (DRAs) are major components of pine (Pinus spp.) oleoresin. They play critical roles in conifer defense against insects and pathogens and as a renewable resource for industrial bioproducts. The core structures of DRAs are formed in secondary (i.e. specialized) metabolism via cycloisomerization of geranylgeranyl diphosphate (GGPP) by diterpene synthases (diTPSs). Previously described gymnosperm diTPSs of DRA biosynthesis are bifunctional enzymes that catalyze the initial bicyclization of GGPP followed by rearrangement of a (+)-copalyl diphosphate intermediate at two discrete class II and class I active sites. In contrast, similar diterpenes of gibberellin primary (i.e. general) metabolism are produced by the consecutive activity of two monofunctional class II and class I diTPSs. Using high-throughput transcriptome sequencing, we discovered 11 diTPS from jack pine (Pinus banksiana) and lodgepole pine (Pinus contorta). Three of these were orthologous to known conifer bifunctional levopimaradiene/abietadiene synthases. Surprisingly, two sets of orthologous PbdiTPSs and PcdiTPSs were monofunctional class I enzymes that lacked functional class II active sites and converted (+)-copalyl diphosphate, but not GGPP, into isopimaradiene and pimaradiene as major products. Diterpene profiles and transcriptome sequences of lodgepole pine and jack pine are consistent with roles for these diTPSs in DRA biosynthesis. The monofunctional class I diTPSs of DRA biosynthesis form a new clade within the gymnosperm-specific TPS-d3 subfamily that evolved from bifunctional diTPS rather than monofunctional enzymes (TPS-c and TPS-e) of gibberellin metabolism. Homology modeling suggested alterations in the class I active site that may have contributed to their functional specialization relative to other conifer diTPSs.Conifer trees, including lodgepole pine (Pinus contorta) and jack pine (Pinus banksiana), produce complex mixtures of mono-, sesqui-, and diterpenoid specialized (i.e. secondary) metabolites, most prominently in the form of oleoresin, that can act as a physical and chemical defense against insect and pathogen attack

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Enzymatic 13C Labeling and Multidimensional NMR Analysis of Miltiradiene Synthesized by Bifunctional Diterpene Cyclase in Selaginella moellendorffii

Cited by 42 publications

References 32 publications

Nonseed plant Selaginella moellendorffii has both seed plant and microbial types of terpene synthases

Nonseed plant Selaginella moellendorffii has both seed plant and microbial types of terpene synthases

Molecular Diversity of Terpene Synthases in the Liverwort Marchantia polymorpha

Evolution of Conifer Diterpene Synthases: Diterpene Resin Acid Biosynthesis in Lodgepole Pine and Jack Pine Involves Monofunctional and Bifunctional Diterpene Synthases

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