(593u) A Purification Method for Thermostable Recombinant Carbonic Anhydrase Proteins Produced in E. Coli

As technologies using thermostable carbonic anhydrases, an enzyme that catalyzes the reversible reaction of carbon dioxide and water to form carbonic acid, are being developed, there is a growing need for an economical, efficient and scalable method for purifying recombinant thermostable carbonic anhydrases from E. coli. We have been developing such a method that purifies thermostable carbonic anhydrases in high yield using only heat and ammonium sulfate. We have used this purification method to produce variants of the carbonic anhydrase from Methanosarcina thermophilia with tags to be used in technologies that require the immobilization of the enzyme. Currently we are investigating the effects of various types of covalent immobilization on the activity and stability of these enzymes. The applications of immobilized carbonic anhydrases include their use in carbon capture and storage technologies. The development of a large-scale purification method for these enzymes is critical for the viability of these technologies.