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Two functionally connected genes encoding a glucosyl-3-phosphoglycerate synthase (GpgS) and an unknown glycosyltransferase (gene Pmob_1143), which we functionally characterized as a mannosylglucosyl-3-phosphoglycerate synthase and designated MggA, were identified in the genome of<jats:italic>Ptg. mobilis<\/jats:italic>. This enzyme used the product of GpgS, glucosyl-3-phosphoglycerate (GPG), as well as GDP-mannose to produce mannosylglucosyl-3-phosphoglycerate (MGPG), the phosphorylated precursor of MGG. The MGPG dephosphorylation was determined in cell extracts, and the native enzyme was partially purified and characterized. Surprisingly, a gene encoding a putative glucosylglycerate synthase (Ggs) was also identified in the genome of<jats:italic>Ptg. mobilis<\/jats:italic>, and an active Ggs capable of producing glucosylglycerate (GG) from ADP-glucose and<jats:sc>d<\/jats:sc>-glycerate was detected in cell extracts and the recombinant enzyme was characterized, as well. Since GG has never been identified in this organism nor was it a substrate for the MggA, we anticipated the existence of a nonphosphorylating pathway for MGG synthesis. We putatively identified the corresponding gene, whose product had some sequence homology with MggA, but it was not possible to recombinantly express a functional enzyme from<jats:italic>Ptg. mobilis<\/jats:italic>, which we named mannosylglucosylglycerate synthase (MggS). In turn, a homologous gene from<jats:italic>Thermotoga maritima<\/jats:italic>was successfully expressed, and the synthesis of MGG was confirmed from GDP-mannose and GG. Based on the measurements of the relevant enzyme activities in cell extracts and on the functional characterization of the key enzymes, we propose two alternative pathways for the synthesis of the rare compatible solute MGG in<jats:italic>Ptg. mobilis<\/jats:italic>.<\/jats:p>","DOI":"10.1128\/jb.01424-09","type":"journal-article","created":{"date-parts":[[2010,1,9]],"date-time":"2010-01-09T02:57:25Z","timestamp":1263005845000},"page":"1624-1633","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":16,"title":["Two Alternative Pathways for the Synthesis of the Rare Compatible Solute Mannosylglucosylglycerate in<i>Petrotoga mobilis<\/i>"],"prefix":"10.1128","volume":"192","author":[{"given":"Chantal","family":"Fernandes","sequence":"first","affiliation":[{"name":"Centro de Neurocie\u0302ncias e Biologia Celular, Universidade de Coimbra, 3004-517 Coimbra, Portugal"}]},{"given":"Vitor","family":"Mendes","sequence":"additional","affiliation":[{"name":"Centro de Neurocie\u0302ncias e Biologia Celular, Universidade de Coimbra, 3004-517 Coimbra, Portugal"}]},{"given":"Joana","family":"Costa","sequence":"additional","affiliation":[{"name":"Centro de Neurocie\u0302ncias e Biologia Celular, Universidade de Coimbra, 3004-517 Coimbra, Portugal"}]},{"given":"Nuno","family":"Empadinhas","sequence":"additional","affiliation":[{"name":"Centro de Neurocie\u0302ncias e Biologia Celular, Universidade de Coimbra, 3004-517 Coimbra, Portugal"},{"name":"Departamento de Cie\u0302ncias da Vida, Universidade de Coimbra, 3001-401 Coimbra, Portugal"}]},{"given":"Carla","family":"Jorge","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qui\u0301mica e Biolo\u0301gica, Universidade Nova de Lisboa, 2780-156 Oeiras, Portugal"}]},{"given":"Pedro","family":"Lamosa","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qui\u0301mica e Biolo\u0301gica, Universidade Nova de Lisboa, 2780-156 Oeiras, Portugal"},{"name":"Centro de Ressona\u0302ncia Magne\u0301tica Anto\u0301nio Xavier, Instituto de Tecnologia Qui\u0301mica e Biolo\u0301gica, Universidade Nova de Lisboa, 2781-901 Oeiras, Portugal"}]},{"given":"Helena","family":"Santos","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qui\u0301mica e Biolo\u0301gica, Universidade Nova de Lisboa, 2780-156 Oeiras, Portugal"}]},{"given":"Milton S.","family":"da Costa","sequence":"additional","affiliation":[{"name":"Departamento de Cie\u0302ncias da Vida, Universidade de Coimbra, 3001-401 Coimbra, Portugal"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/0076-6879(66)08014-5","volume":"8","year":"1966","unstructured":"Ames, B. 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