Organic solutes in the deepest phylogenetic branches of the Bacteria: identification of α(1–6)glucosyl-α(1–2)glucosylglycerate in Persephonella marina
The accumulation of organic solutes was investigated in the thermophilic bacteria Persephonella marina and Marinitoga piezophila, two representatives of the deepest lineages in the domain Bacteria. These organisms grow optimally at around 70 °C in medium containing 3 % NaCl. A new disaccharide, accumulating in Persephonella marina, was identified as α(1–6)glucosyl-α(1–2)glucosylglycerate (GGG), by nuclear magnetic resonance. This identification was validated by comparison with the spectra of the compound obtained by chemical synthesis. Besides GGG, the solute pool of Persephonella marina comprised β-glutamate, di-myo-inositol-1,3′-phosphate and 2-O-α-glucosylglycerate. In contrast, amino acids such as α-glutamate, proline and alanine were the dominant components of the solute pool of Marinitoga piezophila and sugar derivatives were absent. The ability of GGG to protect protein structure against heat denaturation was assessed using model proteins. A genomic search for the biosynthetic pathways of known ionic solutes in Aquificales and Thermotogales shows the inability of this analysis to predict the nature of compatible solutes and underlines the need for efficient cultivation techniques.
KeywordsCompatible solutes Persephonellamarina Thermophiles Glucosylglycerate Glucosylglucosylglycerate
This work was supported by the European Commission, 6th Framework Programme contract COOP-CT-2003-508644, PRODEP and POCI, Portugal (PTDC/BIO/70806/2006) (POCI/V.5/A0004/2005). Technical assistance by Ana I Mingote is acknowledged. The NMR spectrometers are part of The National NMR Network (REDE/1517/RMN/2005), supported by “Programa Operacional Ciência e Inovação (POCTI) 2010” and Fundação para a Ciência e a Tecnologia (FCT). M.V.R. and L.G.G. received fellowships from FCT (SFRH/BPD/80219/2011 and SFRH/BPD/26905/2006). Data for the amino acid analysis was obtained by the Analytical Laboratory, Analytical Services Unit, Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa.
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