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The plant Selaginella moellendorffii possesses enzymes for synthesis and hydrolysis of the compatible solutes mannosylglycerate and glucosylglycerate

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Abstract

A mannosylglycerate synthase (MgS) gene detected in the genome of Selaginella moellendorffii was expressed in E. coli and the recombinant enzyme was purified and characterized. A remarkable and unprecedented feature of this enzyme was the ability to efficiently synthesize mannosylglycerate (MG) and glucosylglycerate (GG) alike, with maximal activity at 50 °C, pH 8.0 and with Mg2+ as reaction enhancer. We have also identified a novel glycoside hydrolase gene in this plant’s genome, which was functionally confirmed to be highly specific for the hydrolysis of MG and GG and named MG hydrolase (MgH), due to its homology with bacterial MgHs. The recombinant enzyme was maximally active at 40 °C and at pH 6.0–6.5. The activity was independent of cations, but Mn2+ was a strong stimulator. Regardless of these efficient enzymatic resources we could not detect MG or GG in S. moellendorffii or in the extracts of five additional Selaginella species. Herein, we describe the properties of the first eukaryotic enzymes for the synthesis and hydrolysis of the compatible solutes, MG and GG.

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Abbreviations

GG:

Glucosylglycerate

GGG:

Glucosylglucosylglycerate

GgS:

Glucosylglycerate synthase

GpgP:

Glucosyl-3-phosphoglycerate phosphatase

GpgS:

Glucosyl-3-phosphoglycerate synthase

MG:

Mannosylglycerate

MGA:

Mannosylglyceramide

MGG:

Mannosylglucosylglycerate

MgH:

Mannosylglycerate hydrolase

MgS:

Mannosylglycerate synthase

MpgP:

Mannosyl-3-phosphoglycerate phosphatase

MpgS:

Mannosyl-3-phosphoglycerate synthase

NMR:

Nuclear magnetic resonance

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

TLC:

Thin layer chromatography

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Acknowledgments

The NMR spectrometers are part of The National NMR Network (REDE/1517/RMN/2005), supported by “Programa Operacional Ciência e Inovação (POCI) 2010” and Fundação para a Ciência e Tecnologia (FCT). This work was supported by FCT, Portugal: projects PTDC/BIA-MIC/105247/2008 and PTDC/BIA-BCM/112459/2009. A. N. acknowledges a scholarship from FCT (SFRH/BD/28907/2006). We wish to thank Joana Costa (Life Sciences Department and CNC, University of Coimbra, Portugal) for advice on the phylogenetic analyses.

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Authors and Affiliations

  1. CNC, Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517, Coimbra, Portugal

    Ana Nobre, Nuno Empadinhas, Maria Fernanda Nobre & Milton S. da Costa

  2. Department of Life Sciences, University of Coimbra, 3001-401, Coimbra, Portugal

    Nuno Empadinhas, Maria Fernanda Nobre & Milton S. da Costa

  3. ITQB, Institute of Chemical and Biological Technology, New University of Lisbon, 2780-901, Oeiras, Portugal

    Eva Correia Lourenço, Christopher Maycock, Maria Rita Ventura & Ana Mingote

  4. Department of Chemistry and Biochemistry, Faculty of Sciences, University of Lisbon, 1749-016, Lisbon, Portugal

    Christopher Maycock

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  1. Ana Nobre
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  2. Nuno Empadinhas
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  3. Maria Fernanda Nobre
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Correspondence to Milton S. da Costa.

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Nobre, A., Empadinhas, N., Nobre, M.F. et al. The plant Selaginella moellendorffii possesses enzymes for synthesis and hydrolysis of the compatible solutes mannosylglycerate and glucosylglycerate. Planta 237, 891–901 (2013). https://doi.org/10.1007/s00425-012-1808-6

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