3V3G

Kinetic and structural studies of thermostabilized mutants of HCA II.

Summary for 3V3G

• Entry DOI: 10.2210/pdb3v3g/pdb
• Related: 3V3F 3V3H 3V3I 3V3J
• Descriptor: Carbonic anhydrase 2, ZINC ION, CHLORIDE ION, ... (4 entities in total)
• Functional Keywords: thermostabile, lyase
• Biological source: Homo sapiens (human)
• Cellular location: Cytoplasm : P00918
• Total number of polymer chains: 1
• Total formula weight: 29356.79

Authors

Boone, C.D.,Fisher, S.Z.,McKenna, R. (deposition date: 2011-12-13, release date: 2012-06-20, Last modification date: 2024-02-28)

Primary citation

Fisher, Z.,Boone, C.D.,Biswas, S.M.,Venkatakrishnan, B.,Aggarwal, M.,Tu, C.,Agbandje-McKenna, M.,Silverman, D.,McKenna, R.
Kinetic and structural characterization of thermostabilized mutants of human carbonic anhydrase II.
Protein Eng.Des.Sel., 25:347-355, 2012

PubMed Abstract: 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.

PubMed: 22691706
DOI: 10.1093/protein/gzs027

Experimental method

X-RAY DIFFRACTION (1.5581 Å)