Expression, reconstruction and characterization of codon-optimized carbonic anhydrase from Hahella chejuensis for CO₂ sequestration application

The high production of functional carbonic anhydrase (CA) is required for practical CO₂ sequestration application mediated by CA. Here, the synthetic gene based on Escherichia coli codon usage of new α-type CA (HC-aCA) of Hahella chejuensis, a Korea marine microorganism, was highly expressed in E. coli. We obtained a high yield of functional HC-aCA by denaturing/refolding process and incorporating zinc ion into its active site. The refolded HC-aCA displayed a half-deactivation temperature of 60 C with maximal activity at 50 C, and had high pH stability in alkali condition with maximal activity at pH 10.0. The esterase activity of HC-aCA almost doubled at high salt concentration ranging from 0.67 to 2.0 M NaCl. HC-aCA catalyzed the conversion of CO₂ to CaCO₃ as calcites form in the presence of Ca²⁺. The refolded HC-aCA could be a promising candidate for the development of efficient CA-based CO₂ sequestration processes.