Abstract
The gammaproteobacterium Thiomicrospira crunogena XCL-2 is an aerobic sulfur-oxidizing hydrothermal vent chemolithoautotroph that has a CO2 concentrating mechanism (CCM), which generates intracellular dissolved inorganic carbon (DIC) concentrations much higher than extracellular, thereby providing substrate for carbon fixation at sufficient rate. This CCM presumably requires at least one active DIC transporter to generate the elevated intracellular concentrations of DIC measured in this organism. In this study, the half-saturation constant (K CO2) for purified carboxysomal RubisCO was measured (276 ± 18 µM) which was much greater than the K CO2 of whole cells (1.03 µM), highlighting the degree to which the CCM facilitates CO2 fixation under low CO2 conditions. To clarify the bioenergetics powering active DIC uptake, cells were incubated in the presence of inhibitors targeting ATP synthesis (DCCD) or proton potential (CCCP). Incubations with each of these inhibitors resulted in diminished intracellular ATP, DIC, and fixed carbon, despite an absence of an inhibitory effect on proton potential in the DCCD-incubated cells. Electron transport complexes NADH dehydrogenase and the bc 1 complex were found to be insensitive to DCCD, suggesting that ATP synthase was the primary target of DCCD. Given the correlation of DIC uptake to the intracellular ATP concentration, the ABC transporter genes were targeted by qRT-PCR, but were not upregulated under low-DIC conditions. As the T. crunogena genome does not include orthologs of any genes encoding known DIC uptake systems, these data suggest that a novel, yet to be identified, ATP- and proton potential-dependent DIC transporter is active in this bacterium. This transporter serves to facilitate growth by T. crunogena and other Thiomicrospiras in the many habitats where they are found.
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Acknowledgments
We are tremendously thankful to the National Science Foundation for their support of this project (NSF-MCB-0643713 to K.M.S.). We would also like to thank Gordon Cannon and Sabine Heinhorst for helpful discussions and for the use of their facilities for carboxysome studies, and the anonymous reviewers for their insightful suggestions, which improved the quality of this manuscript.
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Communicated by Erko Stackebrandt.
USF MCB4404L 2012 are: P. Wanjugi, M. Abdel-Rahim, M. B. Alak, L. J. Astatrjan, V. Bihary, F. Blazekovic, C. Cabrera, G. Camper, T. Chase, J. Dox, A. Echevarria, Q. A. Fisher, C. Georgeades, I. E. Heller, A. N. Hewlett, A. E. Justus, M. Kemp, M. Kondoff, J. P. Martin, E. McClenthan, G. R. Nicolas, J. Paoletti, S. Schuler, M. Skopis, S. R. Subar, E. R. Trebour.
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Menning, K.J., Menon, B.B., Fox, G. et al. Dissolved inorganic carbon uptake in Thiomicrospira crunogena XCL-2 is Δp- and ATP-sensitive and enhances RubisCO-mediated carbon fixation. Arch Microbiol 198, 149–159 (2016). https://doi.org/10.1007/s00203-015-1172-6
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DOI: https://doi.org/10.1007/s00203-015-1172-6