title: Cloning, characterization and anion inhibition study of a β-class carbonic anhydrase from the caries producing pathogen Streptococcus mutans.
abstract: The oral pathogenic bacterium involved in human dental caries formation Streptococcus mutans, encodes for two carbonic anhydrase (CA, EC 4.2.1.1) one belonging to the α- and the other one to the β-class. This last enzyme (SmuCA) has been cloned, characterized and investigated for its inhibition profile with a major class of CA inhibitors, the inorganic anions. Here we show that SmuCA has a good catalytic activity for the CO2 hydration reaction, with kcat 4.2×10(5)s(-1) and kcat/Km of 5.8×10(7)M(-1)×s(-1), being inhibited by cyanate, carbonate, stannate, divannadate and diethyldithiocarbamate in the submillimolar range (KIs of 0.30-0.64mM) and more efficiently by sulfamide, sulfamate, phenylboronic acid and phenylarsonic acid (KIs of 15-46μM). The anion inhibition profile of the S. mutans enzyme is very different from other α- and β-CAs investigated earlier. Identification of effective inhibitors of this new enzyme may lead to pharmacological tools useful for understanding the role of S. mutans CAs in dental caries formation, and eventually the development of pharmacological agents with a new mechanism of antibacterial action.
authors.affiliation: Università degli Studi di Firenze, Laboratorio di Chimica Bioinorganica, Rm. 188, Via della Lastruccia 3, I-50019 Sesto Fiorentino (Firenze), Italy; Department of Chemistry, Faculty of Art & Science, Balikesir University, Balikesir, Turkey.
authors.forename: Nurcan
authors.initials: N
authors.lastname: Dedeoglu
authors.affiliation: Istituto di Biochimica delle Proteine-CNR, Via P. Castellino 111, 80131 Napoli, Italy.
authors.forename: Viviana
authors.initials: V
authors.lastname: De Luca
authors.affiliation: Department of Chemistry, Faculty of Art & Science, Balikesir University, Balikesir, Turkey.
authors.forename: Semra
authors.initials: S
authors.lastname: Isik
authors.affiliation: Department of Biology, Faculty of Art & Science, Balikesir University, Balikesir, Turkey.
authors.forename: Hatice
authors.initials: H
authors.lastname: Yildirim
authors.affiliation: Department of Biology, Faculty of Art & Science, Balikesir University, Balikesir, Turkey.
authors.forename: Feray
authors.initials: F
authors.lastname: Kockar
authors.affiliation: Istituto di Biochimica delle Proteine-CNR, Via P. Castellino 111, 80131 Napoli, Italy.
authors.forename: Clemente
authors.initials: C
authors.lastname: Capasso
authors.affiliation: Università degli Studi di Firenze, Polo Scientifico, Dipartimento NEUROFABA, Sezione di Scienze Farmaceutiche e Nutraceutiche, Via Ugo Schiff 6, 50019 Sesto Fiorentino (Firenze), Italy. Electronic address: claudiu.supuran@unifi.it.
authors.forename: Claudiu T
authors.initials: CT
authors.lastname: Supuran
mesh_terms: D000595:Amino Acid Sequence; D000838:Anions; D001152:Arsenicals / Q000737:chemistry*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D001897:Boronic Acids / Q000737:chemistry*; D002245:Carbon Dioxide / Q000737:chemistry / Q000378:metabolism; D002257:Carbonic Anhydrase Inhibitors / Q000737:chemistry*; D002256:Carbonic Anhydrases / Q000737:chemistry* / Q000235:genetics; D003001:Cloning, Molecular; D003731:Dental Caries / Q000382:microbiology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D013045:Species Specificity; D013295:Streptococcus mutans / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D013451:Sulfonic Acids / Q000737:chemistry*
chemical_list: D000838:Anions; D001152:Arsenicals; D001426:Bacterial Proteins; D001897:Boronic Acids; D002257:Carbonic Anhydrase Inhibitors; D011994:Recombinant Proteins; D013451:Sulfonic Acids; D002245:Carbon Dioxide; C020979:benzenearsonic acid; C005741:sulfamic acid; D002256:Carbonic Anhydrases; C010686:benzeneboronic acid
keywords: Anion inhibitor; Carbonic anhydrase; Streptococcus mutans; β-Class enzyme