Recombinant Carbonic Anhydrase Variants for High-Temperature Carbon Capture

The patent applies ancestral sequence reconstruction to identify and synthesize ancient CA sequences that inherently possess superior thermostability. By changing the amino acid sequence parameters to match ancestral forms, the enzyme achieves enhanced stability under harsh industrial conditions while maintaining catalytic activity. This is demonstrated by the ancestral CA variants showing improved thermostability compared to modern CAs.

The patent creates composite enzyme structures by combining ancestral sequence elements with modern CA frameworks. The reconstructed ancestral sequences integrate beneficial amino acid residues from multiple modern CA variants, creating a composite enzyme that combines the thermostability of ancient enzymes with the catalytic efficiency of modern variants.

The patent systematically varies amino acid sequences by reconstructing ancestral forms and introducing targeted mutations. By changing specific sequence parameters while maintaining the overall fold and active site architecture, the enzyme achieves both enhanced thermostability and preserved catalytic activity. The ancestral sequences naturally balance stability and activity parameters.

The patent creates enzyme variants with dynamic stability properties, allowing the enzyme to maintain its structure at high temperatures while preserving the flexibility needed for catalysis. The ancestral CA variants exhibit dynamic behavior that enables them to remain stable under harsh conditions while maintaining catalytic competence, as evidenced by their performance in CO2 hydration reactions at elevated temperatures.