Characteristics of recombinant α-carbonic anhydrase of polyextremophilic bacterium Bacillus halodurans TSLV1

•First report on heterologous production of α-carbonic anhydrase from Bacillus halodurans TSLV1 (BhCA) in E. coli.•rBhCA is a novel alkalistable carbonic anhydrase active in the pH range of 6–11.•rBhCA is moderately thermostable with stability in the presence of SOx and NOx.•rBhCA is stimulated by sulphate and displays higher thermostability in its presence.•rBhCA is very efficient in accelerating mineralization of CO2 converting it to calcite form.•rBhCA can function as alkalistable virtual peroxidase upon substitution of active site Zn2+ with Mn2+.

Carbonic anhydrase (CA) is a biocatalyst that catalyzes the hydration of CO2 to bicarbonate and protons, thus useful in mitigating green house effect by sequestering CO2 from various point sources. An alkalistable and moderately thermostable α- carbonic anhydrase encoding gene (BhCA) from Bacillus halodurans TSLV1 has been cloned and expressed in Escherichia coli. A 31.4-fold enhancement in CA production was achieved due to cloning and expression in E. coli. About 50% of the CA produced was secreted when recombinant E. coli with BhCA-pET22b was cultivated in a medium with EDTA and lysozyme because of the efficient pelB leader sequence. rBhCA is a ∼75 kDa homodimeric protein with a Tm of 72 °C and T1/2 values of 66 and 24 min at 50 and 60 °C, respectively. SDM analysis revealed that H137, H139, H156 and H110 present in the active site play an important role in catalysis. Mineralization of CO2 using rBhCA led to the accelerated precipitation of CaCO3 in calcite form. rBhCA also functions as an efficient virtual peroxidase when Zn2+ is substituted with Mn2+.