Abstract
Almost all the inorganic carbon on Earth is converted into biomass via the Calvin–Benson–Bassham (CBB) cycle. Here, the central carboxylation reaction is catalyzed by ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO), which can be found in numerous primary producers including plants, algae, cyanobacteria, and many autotrophic bacteria. Although RubisCO possesses a crucial role in global biomass production, it is not a perfect catalyst. Therefore, research interest persists on accessing the full potential of yet unexplored RubisCOs. We recently developed an activity-based screen suited to seek active recombinant RubisCOs from the environment—independent of the native host’s culturability. Here, we applied this screen to twenty pre-selected genomic fosmid clones from six cultured proteobacteria to demonstrate that a broad range of phylogenetically distinct RubisCOs can be targeted. We then screened 12,500 metagenomic fosmid clones from six distinct hydrothermal vents and identified forty active RubisCOs. Additional sequence-based screening uncovered eight further RubisCOs, which could then also be detected by a modified version of the screen. Seven were active form III RubisCOs from yet uncultured Archaea. This indicates the potential of the activity-based screen to detect RubisCO enzymes even from organisms that would not be expected to be targeted.
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Acknowledgements
We thank the captain and crews of the RV Meteor, the Maria S. Merian as well as the ROV Kiel6000 (GEOMAR, Kiel) for helping us to obtain deep-sea vent samples. We thank Wenke Bahnsen, Dagmar Svensson, Anna Ulatowski, and Nicole Adam for excellent technical assistance in the laboratory. The work was supported by grants from the Deutsche Forschungsgemeinschaft (DFG) (PE1549/5-1) and the DFG priority program 1144 “From Mantle to Ocean: Energy-, Material- and Life-cycles at Spreading Axes”.
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Böhnke, S., Perner, M. Seeking active RubisCOs from the currently uncultured microbial majority colonizing deep-sea hydrothermal vent environments. ISME J 13, 2475–2488 (2019). https://doi.org/10.1038/s41396-019-0439-3
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DOI: https://doi.org/10.1038/s41396-019-0439-3
