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Carbonic anhydrase enzymes catalyse the reversible hydration of carbon dioxide to bicarbonate. A thermophilic Thermovibrio ammonificans α-carbonic anhydrase (TaCA) has been expressed in Escherichia coli and structurally and biochemically characterized. The crystal structure of TaCA has been determined in its native form and in two complexes with bound inhibitors. The tetrameric enzyme is stabilized by a unique core in the centre of the molecule formed by two intersubunit disulfides and a single lysine residue from each monomer that is involved in intersubunit ionic interactions. The structure of this core protects the intersubunit disulfides from reduction, whereas the conserved intrasubunit disulfides are not formed in the reducing environment of the E. coli host cytosol. When oxidized to mimic the environment of the periplasmic space, TaCA has increased thermostability, retaining 90% activity after incubation at 70°C for 1 h, making it a good candidate for industrial carbon-dioxide capture. The reduction of all TaCA cysteines resulted in dissociation of the tetrameric molecule into monomers with lower activity and reduced thermostability. Unlike other characterized α-carbonic anhydrases, TaCA does not display esterase activity towards p-nitrophenyl acetate, which appears to result from the increased rigidity of its protein scaffold.

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Visualization experiments using one-dimensional (1-D) porous media made of Plexiglas and two-dimensional glass porous networks were conducted to obtain qualitative and quantitative information concerning the precipitation and crystal growth of CaCO3 under varying flow and concentration conditions. Supersaturated solutions were prepared by mixing sodium bicarbonate and calcium chloride solutions before the pore networks. Nucleation and crystal growth were assumed to occur within the porous media. Changes in the initial and final solution composition were monitored. At low initial supersaturation values (SRinitial), a few crystals were observed within the flow channels and crystal growth took place exclusively on the newly formed crystals. As the SRinitial increased, more crystals were formed along the flow channels and new crystallites were continuously formed during the course of the experiments. Nucleation and crystal growth were not uniform. The crystal growth rates depended on the initial value of SR and flow path inside the medium. Porosity for the 2-D networks decreased when the SRinitial was high or when calcite-cemented sand was used as substrate.