A new type of carboxysomal carbonic anhydrase in sulfur chemolithoautotrophs from alkaline environments.

UNLABELLED

Autotrophic bacteria are able to fix CO in a great diversity of habitats, even though this dissolved gas is relatively scarce at neutral pH and above. As many of these bacteria rely on CO fixation by ribulose 1,5-bisphospate carboxylase/oxygenase (RubisCO) for biomass generation, they must compensate for the catalytical constraints of this enzyme with CO-concentrating mechanisms (CCMs). CCMs consist of CO and HCO transporters and carboxysomes. Carboxysomes encapsulate RubisCO and carbonic anhydrase (CA) within a protein shell and are essential for the operation of a CCM in autotrophic that use the Calvin-Benson-Basham cycle. Members of the genus lack genes homologous to those encoding previously described CA, and prior to this work, the mechanism of function for their carboxysomes was unclear. In this paper, we provide evidence that a member of the recently discovered iota family of carbonic anhydrase enzymes (ιCA) plays a role in CO fixation by carboxysomes from members of and potentially other . Carboxysome enrichments from and were found to have CA activity and contain ιCA, which is encoded in their carboxysome loci. When the gene encoding ιCA was interrupted in , cells could no longer grow under low-CO conditions, and CA activity was no longer detectable in their carboxysomes. When ιCA was expressed in a strain of lacking native CA activity, this strain recovered an ability to grow under low CO conditions, and CA activity was present in crude cell extracts prepared from this strain.

IMPORTANCE

Here, we provide evidence that iota carbonic anhydrase (ιCA) plays a role in CO fixation by some organisms with CO-concentrating mechanisms; this is the first time that ιCA has been detected in carboxysomes. While ιCA genes have been previously described in other members of bacteria, this is the first description of a physiological role for this type of carbonic anhydrase in this domain. Given its distribution in alkaliphilic autotrophic bacteria, ιCA may provide an advantage to organisms growing at high pH values and could be helpful for engineering autotrophic organisms to synthesize compounds of industrial interest under alkaline conditions.