Kinetic and structural characterization of thermostabilized mutants of human carbonic anhydrase II.


Carbonic anhydrases (CAs) are ubiquitous enzymes that catalyze the reversible hydration/dehydration of carbon dioxide/bicarbonate. As such, there is enormous industrial interest in using CA as a bio-catalyst for carbon sequestration and biofuel production. However, to ensure cost-effective use of the enzyme under harsh industrial conditions, studies were initiated to produce variants with enhanced thermostability while retaining high solubility and catalytic activity. Kinetic and structural studies were conducted to determine the structural and functional effects of these mutations. X-ray crystallography revealed that a gain in surface hydrogen bonding contributes to stability while retaining proper active site geometry and electrostatics to sustain catalytic efficiency. The kinetic profiles determined under a variety of conditions show that the surface mutations did not negatively impact the carbon dioxide hydration or proton transfer activity of the enzyme. Together these results show that it is possible to enhance the thermal stability of human carbonic anhydrase II by specific replacements of surface hydrophobic residues of the enzyme. In addition, combining these stabilizing mutations with strategic active site changes have resulted in thermostable mutants with desirable kinetic properties.

Submission Details

ID: FjiZCxqr3

Submitter: Shu-Ching Ou

Submission Date: Dec. 5, 2018, 3:39 p.m.

Version: 1

Publication Details
Fisher Z;Boone CD;Biswas SM;Venkatakrishnan B;Aggarwal M;Tu C;Agbandje-McKenna M;Silverman D;McKenna R,Protein Eng Des Sel (2012) Kinetic and structural characterization of thermostabilized mutants of human carbonic anhydrase II. PMID:22691706
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Carbonic anhydrase 2 P00918 CAH2_HUMAN