Low Rubisco activity was detected that could not account for the carbon dioxide (CO2) fixation rate; in addition, phosphoribulokinase activity was not found. The generation of ribulose 1,5-bisphosphate from 5-phospho-d-ribose 1-pyrophosphate was observed, but not from AMP; these sources for ribulose 1,5-bisphosphate have been proposed before. Our data indicate that the reductive acetyl-CoA pathway is the only functioning CO2 fixation pathway in ‘A.

lithotrophicus’. To date, six autotrophic carbon dioxide (CO2) fixation pathways have been found in nature, three of which buy Ku-0059436 occur in Archaea (Berg et al., 2010a). Whereas Crenarchaeota use either the dicarboxylate/hydroxybutyrate or the hydroxypropionate/hydroxybutyrate cycle (Fig. 1), all autotrophic Euryarchaeota studied so far use the reductive acetyl-CoA pathway (Fig. 1c) (Berg et al., 2010a). The functioning of this pathway in Euryarchaeota conforms to the fact that most autotrophic Euryarchaeota are methanogens. They use much of the enzymes involved in energy generation during methanogenesis also for autotrophic acetyl-CoA synthesis. The only known exceptions to

this rule are members of Archaeoglobi (Huber & Stetter, 2001) and perhaps Ferroplasma acidiphilum (Golyshina et al., 2000), whose ability to grow autotrophically was questioned (Dopson et al., 2004). Representatives of the class Archaeoglobi (with only one order and one family, Archaeoglobales and Archaeoglobaceae) are hyperthermophiles Osimertinib that grow by means

of anaerobic respiration by oxidizing organic substrates or molecular hydrogen (in some cases, also Fe2+ or S2−) (Huber & Stetter, 2001). The Archaeoglobaceae comprise three Thiamet G genera: Archaeoglobus, Ferroglobus and Geoglobus. Besides Archaeoglobus profundus and Archaeoglobus infectus, all species can grow autotrophically, with ‘Archaeoglobus lithotrophicus’ being an obligate autotroph (Stetter et al., 1993). Biochemical studies revealed the presence of the enzymes of the reductive acetyl-CoA pathway in ‘A. lithotrophicus’ and Ferroglobus placidus (Vorholt et al., 1995, 1997). The corresponding genes also exist in the Archaeoglobus fulgidus genome (Klenk et al., 1997), whereas the genome of the heterotrophic A. profundus lacks the gene for the key enzyme of this pathway, CO-dehydrogenase/acetyl-CoA synthase (von Jan et al., 2010). Therefore, these data suggest that autotrophic Archaeoglobaceae use the reductive acetyl-CoA pathway for CO2 fixation. Interestingly, the genome of A. fulgidus also harbors, besides the genes of the reductive acetyl-CoA pathway, three copies of genes encoding homologues of the 4-hydroxybutyryl-CoA dehydratase. In contrast, this key enzyme of the dicarboxylate/hydroxybutyrate and hydroxypropionate/hydroxybutyrate cycles is absent in the heterotrophic A. profundus.