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Dissolved Inorganic Carbon-Accumulating Complexes from Autotrophic Bacteria from Extreme Environments.

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Novel bacterial complexes effectively increase intracellular dissolved inorganic carbon (DIC) levels, aiding CO2 fixation across diverse species. This DIC accumulation mechanism is conserved in various organisms and may be engineered for industrial applications.

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Area of Science:

  • Microbiology
  • Biochemistry
  • Molecular Biology

Background:

  • Autotrophic organisms utilize mechanisms to concentrate dissolved inorganic carbon (DIC) for CO2 fixation, especially in low-carbon environments.
  • While studied in Cyanobacteria, DIC concentrating mechanisms in other bacterial and archaeal phyla are less understood.
  • Novel multisubunit membrane-spanning complexes have been identified, with homologs distributed across 17 phyla of Bacteria and Archaea.

Purpose of the Study:

  • To investigate whether DIC accumulation is a conserved function across diverse multisubunit complexes found in Bacteria and Archaea.
  • To characterize the subunit requirements and substrate specificity of these novel DIC-accumulating complexes.
  • To assess the potential for heterologous expression and application of these complexes in engineered systems.

Main Methods:

  • Seven representative multisubunit complexes from diverse phyla and subunit configurations were selected for study.
  • Heterologous expression of these complexes was performed in a high-CO2-requiring, carbonic anhydrase-deficient Escherichia coli strain.
  • Growth rescue under low-CO2 conditions, intracellular DIC measurements, isotopic disequilibrium experiments, and ionophore treatments were employed for characterization.

Main Results:

  • All seven expressed complexes rescued the low-CO2 growth defect of the E. coli host.
  • Six of the seven complexes demonstrably elevated intracellular DIC concentrations.
  • Two- and three-subunit complexes required all subunits for function, utilized CO2 as a substrate, and appeared to couple proton potential to DIC accumulation.

Conclusions:

  • DIC accumulation is a conserved trait among diverse multisubunit complexes from various bacterial and archaeal phyla.
  • These complexes exhibit functional conservation across different organisms and habitats, suggesting a universal role in carbon acquisition.
  • Successful heterologous expression in E. coli highlights their potential for engineering applications in CO2 utilization and biomass synthesis.