Scaling down recombinant carbonic anhydrase isolation with immobilized metal ion chromatography (IMAC): Harnessing enzymatic carbon dioxide capture and mineralization

•CA purification was optimized using IMAC.•AEA-COOH-CA membranes enhanced CA catalytic capabilities.•AE-COOH-CA membranes facilitated CO2 conversion and CaCO3 formation.•AEA-COOH-CA membranes exhibited 473.42 WAU/g-membrane with CaCO₃ precipitation of 83.90 mg. Human activities have led to increased atmospheric CO2 levels, raising concerns about climate change. Carbonic anhydrase (CA) enzymes show promise for transforming CO2 into valuable products like calcium carbonate (CaCO3) through mineralization. Purifying and immobilizing CA enzymes on nanofiber membranes enhances their catalytic activity, enabling efficient CO2 conversion and mineralization. Recombinant CA was purified using immobilized metal affinity chromatography (IMAC), optimizing pH, biomass concentration, flow rate, and loading volume for maximum efficiency. The CA enzyme was then immobilized onto a weak ion exchange nanofiber membrane functionalized with AEA-COOH to create the CA-modified membrane (AEA-COOH-CA), enhancing CO2 conversion and CaCO3 mineralization. Optimal purification conditions (pH 7, 1 % biomass, 0.1 mL/min flow rate, 1.0 mL loading volume) were determined using IMAC. The CA-modified membrane effectively converted CO2 and mineralized CaCO3, demonstrating the potential for environmental CO2 sequestration. The immobilized CA activities of the AEA-COOH-CA nanofiber membranes exhibited 473.42 WAU/g-membrane, corresponding to 7.10 WAU per membrane piece. The CaCO3 precipitation reached 83.90 mg, with a precipitation efficiency of 11.82 mg CaCO3/WAU. These findings underscore the promise of enzymatic carbon capture using CA-modified membranes, offering a sustainable solution for greenhouse gas mitigation. [Display omitted]