title: Biochemical properties of a new α-carbonic anhydrase from the human pathogenic bacterium, Vibrio cholerae.
abstract: Abstract Vibrio cholerae, a Gram-negative bacterium, is the causative agent of cholera and colonizes the upper small intestine where sodium bicarbonate is present at a high concentration. Sodium bicarbonate is a potential inducer of virulence gene expression. Bacteria can increase cytosolic bicarbonate levels through the existence of transporter family proteins or through the action of metalloenzymes, called carbonic anhydrases (CAs, EC 4.2.1.1). Vibrio cholerae, lacking of transporter proteins in its genome, utilizes the CA system to accumulate bicarbonate into the cell suggesting a pivotal role of this metalloenzymes in the microbial virulence. Here, we report for the first time the characterization of the α-CA of V. cholerae (VchCA), which has been identified by translated genome inspection. The α-CA encoding gene was cloned and expressed in Escherichia coli and the recombinant protein purified to homogeneity. This investigation aimed to study the biochemical properties of VchCA and to provide preliminary insights in the field of this pathogen virulence. VchCA has a low esterase activity with 4-nitrophenyl acetate as substrate, and a high activity for the hydration of CO2 to bicarbonate.
authors: Istituto di Biochimica delle Proteine , CNR, Napoli , Italy .
authors: Sonia
authors: S
authors: Del Prete
authors: Viviana
authors: V
authors: De Luca
authors: Andrea
authors: A
authors: Scozzafava
authors: Vincenzo
authors: V
authors: Carginale
authors: Claudiu T
authors: CT
authors: Supuran
authors: Clemente
authors: C
authors: Capasso
mesh_terms: D000595:Amino Acid Sequence; D002256:Carbonic Anhydrases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006358:Hot Temperature; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D014734:Vibrio cholerae / Q000201:enzymology*
chemical_list: D002256:Carbonic Anhydrases