The gene for β-class carbonic anhydrase (CA), which was designated as cahB1, was cloned from the genomic library of the alkaliphilic cyanobacterium Microcoleus chthonoplastes. The product of the cahB1 gene was expressed in Escherichia coli. The protein revealed high specific activity of CA, which was inhibited with ethoxyzolamide. The maximum activity of the recombinant CA was detected at alkaline pH (∼9.0) and its minimum - at neutral pH (∼7.0). Western blotting analysis with the antibodies raised against the recombinant CahB1 protein revealed its localization in cell envelopes of M. chthonoplastes. Immunocytochemical localization of the CahB1 in cells confirmed its extracellular location. The newly characterized CahB1 of Microcoleus was similar in amino acid and nucleotide sequences to well known β-CAs of Synechococcus sp. PCC 7942 (IcfA) and Synechocystis sp. PCC 6803 (CcaA), although those CAs were attributed to the carboxysomal shells of cyanobacteria. Previously we have reported β-class CA which was associated with PS II of alkaliphilic cyanobacteria. Here we first report extracellular localization of β-class CA and provide a scheme for its possible involvement in the maintenance of a balance between external sources of inorganic carbon and photosynthesis in extreme environments of soda lakes.
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