Biochemical properties of a novel and highly thermostable bacterial α-carbonic anhydrase from Sulfurihydrogenibium yellowstonense YO3AOP1.
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.
CNR, Istituto di Biochimica delle Proteine (IBP), Napoli, Italy. c.capasso@ibp.cnr.it
Clemente
C
Capasso
Viviana
V
De Luca
Vincenzo
V
Carginale
Raffaele
R
Cannio
Mosè
M
Rossi
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002245:Carbon Dioxide / Q000737:chemistry*; D024402:Carbonic Anhydrase II / Q000737:chemistry; D002256:Carbonic Anhydrases / Q000737:chemistry* / Q000302:isolation & purification; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D057075:Enzyme Assays; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D006091:Gram-Negative Chemolithotrophic Bacteria / Q000737:chemistry* / Q000201:enzymology; D006358:Hot Temperature; D055767:Immobilized Proteins / Q000737:chemistry / Q000302:isolation & purification; D007700:Kinetics; D009596:Nitrophenols / Q000737:chemistry*; D011140:Polyurethanes; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D013379:Substrate Specificity
D001426:Bacterial Proteins; D055767:Immobilized Proteins; D009596:Nitrophenols; D011140:Polyurethanes; D011994:Recombinant Proteins; D002245:Carbon Dioxide; C008642:4-nitrophenyl acetate; C028279:polyurethane foam; D024402:Carbonic Anhydrase II; D002256:Carbonic Anhydrases