Biochemical properties of a novel and highly thermostable bacterial α-carbonic anhydrase from Sulfurihydrogenibium yellowstonense YO3AOP1

J Enzyme Inhib Med Chem. 2012 Dec;27(6):892-7. doi: 10.3109/14756366.2012.703185. Epub 2012 Jul 18.

Abstract

A new carbonic anhydrase (CA, EC 4.2.1.1) from the thermophilic bacterium Sulfurihydrogenibium yellowstonense YO3AOP1 was identified and characterized. The bacterial carbonic anhydrase gene was expressed in Escherichia coli yielding an active enzyme, which was purified in large amounts. The recombinant protein (SspCA) was found to belong to the α-CA class and displays esterase activity. The kinetic parameters were determined by using CO(2) and p-nitrophenylacetate (p-NpA) as substrates. The bacterial enzyme presented specific activity comparable to that of bovine carbonic anhydrase (bCA II) but it showed biochemical properties never observed for the mammalian enzyme. The thermophilic enzyme, in fact, was endowed with high thermostability and with unaltered residual activity after prolonged exposure to heat up to 100°C. SspCA and the bovine carbonic anhydrase (bCA II) were immobilized within a polyurethane (PU) foam. The immobilized bacterial enzyme was found to be active and stable at 100°C up to 50 h.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / isolation & purification
  • Carbon Dioxide / chemistry*
  • Carbonic Anhydrase II / chemistry
  • Carbonic Anhydrases / chemistry*
  • Carbonic Anhydrases / isolation & purification
  • Cattle
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Assays
  • Enzyme Stability
  • Escherichia coli / genetics
  • Gram-Negative Chemolithotrophic Bacteria / chemistry*
  • Gram-Negative Chemolithotrophic Bacteria / enzymology
  • Hot Temperature
  • Immobilized Proteins / chemistry
  • Immobilized Proteins / isolation & purification
  • Kinetics
  • Nitrophenols / chemistry*
  • Polyurethanes
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Substrate Specificity

Substances

  • Bacterial Proteins
  • Immobilized Proteins
  • Nitrophenols
  • Polyurethanes
  • Recombinant Proteins
  • Carbon Dioxide
  • 4-nitrophenyl acetate
  • polyurethane foam
  • Carbonic Anhydrase II
  • Carbonic Anhydrases