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Evidence for Autotrophic CO₂ Fixation via the Reductive Tricarboxylic Acid Cycle by Members of the Subdivision of Proteobacteria

Biology Department, Woods Hole Oceanographic Institution, Woods Hole, Massachusetts 02543,¹ Mikrobiologie, Institut für Biologie II, Universität Freiburg, Schänzlestrasse 1, 79104 Freiburg, Germany,² NASA Astrobiology Institute, Marine Biological Laboratory, Woods Hole, Massachusetts 02543³

Received 15 September 2004/ Accepted 17 January 2005

Based on 16S rRNA gene surveys, bacteria of the subdivision of proteobacteria have been identified to be important members of microbial communities in a variety of environments, and quite a few have been demonstrated to grow autotrophically. However, no information exists on what pathway of autotrophic carbon fixation these bacteria might use. In this study, Thiomicrospira denitrificans and Candidatus Arcobacter sulfidicus, two chemolithoautotrophic sulfur oxidizers of the subdivision of proteobacteria, were examined for activities of the key enzymes of the known autotrophic CO₂ fixation pathways. Both organisms contained activities of the key enzymes of the reductive tricarboxylic acid cycle, ATP citrate lyase, 2-oxoglutarate:ferredoxin oxidoreductase, and pyruvate:ferredoxin oxidoreductase. Furthermore, no activities of key enzymes of other CO₂ fixation pathways, such as the Calvin cycle, the reductive acetyl coenzyme A pathway, and the 3-hydroxypropionate cycle, could be detected. In addition to the key enzymes, the activities of the other enzymes involved in the reductive tricarboxylic acid cycle could be measured. Sections of the genes encoding the - and ß-subunits of ATP citrate lyase could be amplified from both organisms. These findings represent the first direct evidence for the operation of the reductive tricarboxylic acid cycle for autotrophic CO₂ fixation in -proteobacteria. Since -proteobacteria closely related to these two organisms are important in many habitats, such as hydrothermal vents, oxic-sulfidic interfaces, or oilfields, these results suggest that autotrophic CO₂ fixation via the reductive tricarboxylic acid cycle might be more important than previously considered.

Contribution number 11288 of the Woods Hole Oceanographic Institution.

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