Saccharomyces cerevisiae beta-carbonic anhydrase: ınhibition and activation studies

Abstract

The beta-carbonic anhydrase from Saccharomyces cerevisiae (CA, EC 4.2.1.1), scCA, which is encoded by the Nce103 gene, is an effective catalyst for CO2 hydration to bicarbonate and protons, with a k(cat) of 9.4 x 10(5) s(-1), and k(cat)/K-M of 9.8 x 10(7) M-1.s(-1). Its inhibition with anions and sulfonamides has been investigated, as well as its activation with amines and amino acids. Bromide, iodide and sulfamide, were the best anion inhibitors, with K(I)s of 8.7 - 10.8 mu M. Benzenesulfonamides substituted in 2-, 4- and 3,4-positions with amino, alkyl, halogeno and hydroxyalkyl moieties had K(I)s in the range of 0.976 - 18.45 mu M. Better inhibition (K(I)s in the range of 154 654 nM) was observed for benzenesulfonamides incorporating aminoalkyl/carboxyalkyl moieties or halogenosulfanilamides; benzene-1,3-disulfonamides; simple heterocyclic sulfonamides and sulfanilyl-sulfonamides. The clinically used sulfonamides/sulfamate (acetazolamide, ethoxzolamide, methazolamide, dorzolamide, topiramate, celecoxib, etc.) generally showed effective scCA inhibitory activity, with K(I)s in the range of 82.6 - 133 nM. The best inhibitor (K-I of 15.1 nM) was 4-(2-amino-pyrimidin-4-yl)-benzenesulfonamide. L-adrenaline and some piperazines incorporating aminoethyl moieties were the most effective scCA activators. These studies may lead to a better understanding of the role of this enzyme in yeasts/fungi, and since the Nce103 gene is also present in many pathogenic organisms (Candida spp., Cryptococcus neoformans, etc) they may be useful to develop antifungal drugs.