Abstract: Global climate change and “carbon peaking and carbon neutrality” targets are driving the development of sustainable CO₂ sequestration technologies. Carbonic anhydrase (CA), a highly efficient metalloenzyme, offers a promising solution. It catalyzes CO₂ hydration to form stable carbonate precipitates for long-term carbon storage. The fundamental properties of CA, involving its structure, catalytic mechanism, and critical performance-affecting factors such as pH, temperature and inhibitors are summarized. Immobilization technology is emphasized as a key strategy to improve its stability and reusability. Furthermore, the review explores diverse applications of CA-mediated mineralization, from concrete repair and soil remediation to enhancing ocean carbon sinks and managing industrial gases. However, significant challenges remain before large-scale implementation. These include high enzyme costs, difficulties in large-scale production, poor long-term stability in the field, and unfavorable process economics. Therefore, future research should focus on screening and engineering highly active and inhibition-resistant CA variants, developing low-cost immobilization carriers with enhanced environmental adaptability, and further improving the regulation of mineral precipitation processes and their spatial characteristics. These efforts are essential to facilitate the stable scale-up of CA-based biomineralization systems under practical application conditions and to enhance their overall applicability.