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Applied and Environmental Microbiology, June 2003, p. 3272-3279, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3272-3279.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

The Bacterium Thermus thermophilus, Like Hyperthermophilic Archaea, Uses a Two-Step Pathway for the Synthesis of Mannosylglycerate

Nuno Empadinhas,1 Luciana Albuquerque,1 Anke Henne,2 Helena Santos,3 and Milton S. da Costa1*

Departamento de Bioquímica and Centro de Neurociências de Coimbra, Universidade de Coimbra, 3004-517 Coimbra,1 Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, 2780-156 Oeiras, Portugal,3 Göttingen Genomics Laboratory, Institut für Mikrobiologie und Genetik, 37077 Göttingen, Germany2

Received 3 February 2003/ Accepted 25 March 2003

The biosynthetic pathway for the synthesis of the compatible solute {alpha}-mannosylglycerate (MG) in the thermophilic bacterium Thermus thermophilus HB27 was identified based on the activities of recombinant mannosyl-3-phosphoglycerate synthase (MPGS) (EC 2.4.1.217) and mannosyl-3-phosphoglycerate phosphatase (MPGP) (EC 3.1.3.70). The sequences of homologous genes from the archaeon Pyrococcus horikoshii were used to identify MPGS and MPGP genes in T. thermophilus HB27 genome. Both genes were separately cloned and overexpressed in Escherichia coli, yielding 3 to 4 mg of pure recombinant protein per liter of culture. The molecular masses were 43.6 and 28.1 kDa for MPGS and MPGP, respectively. The recombinant MPGS catalyzed the synthesis of {alpha}-mannosyl-3-phosphoglycerate (MPG) from GDP-mannose and D-3-phosphoglycerate, while the recombinant MPGP catalyzed the dephosphorylation of MPG to MG. The recombinant MPGS had optimal activity at 80 to 90°C and a pH optimum near 7.0; MPGP had maximal activity between 90 and 95°C and at pH 6.0. The activities of both enzymes were strictly dependent on divalent cations; Mn2+ was most effective for MPGS, while Mn2+, Co2+, Mg2+, and to a lesser extent Ni2+ activated MPGP. The organization of MG biosynthetic genes in T. thermophilus HB27 is different from the P. horikoshii operon-like structure, since the genes involved in the conversion of fructose-6-phosphate to GDP-mannose are not found immediately downstream of the contiguous MPGS and MPGP genes. The biosynthesis of MG in the thermophilic bacterium T. thermophilus HB27, proceeding through a phosphorylated intermediate, is similar to the system found in hyperthermophilic archaea.

* Corresponding author. Mailing address: Departamento de Bioquímica and Centro de Neurociências de Coimbra, Universidade de Coimbra, 3004-517 Coimbra, Portugal. Phone: 351-239 824024. Fax: 351-239 826798. E-mail: milton{at}ci.uc.pt.

Applied and Environmental Microbiology, June 2003, p. 3272-3279, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3272-3279.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Costa, J., Empadinhas, N., da Costa, M. S. (2007). Glucosylglycerate Biosynthesis in the Deepest Lineage of the Bacteria: Characterization of the Thermophilic Proteins GpgS and GpgP from Persephonella marina. J. Bacteriol. 189: 1648-1654 [Abstract] [Full Text]  
  • Costa, J., Empadinhas, N., Goncalves, L., Lamosa, P., Santos, H., da Costa, M. S. (2006). Characterization of the Biosynthetic Pathway of Glucosylglycerate in the Archaeon Methanococcoides burtonii. J. Bacteriol. 188: 1022-1030 [Abstract] [Full Text]  
  • Neves, C., da Costa, M. S., Santos, H. (2005). Compatible Solutes of the Hyperthermophile Palaeococcus ferrophilus: Osmoadaptation and Thermoadaptation in the Order Thermococcales. Appl. Environ. Microbiol. 71: 8091-8098 [Abstract] [Full Text]  
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  • Borges, N., Marugg, J. D., Empadinhas, N., Costa, M. S. d., Santos, H. (2004). Specialized Roles of the Two Pathways for the Synthesis of Mannosylglycerate in Osmoadaptation and Thermoadaptation of Rhodothermus marinus. J. Biol. Chem. 279: 9892-9898 [Abstract] [Full Text]  


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