The Effect of Substituted Benzene-Sulfonamides and Clinically Licensed Drugs on the Catalytic Activity of CynT2, a Carbonic Anhydrase Crucial for Escherichia coli Life Cycle

Abstract

Proteins are relevant antimicrobial drug targets, and among them, enzymes represent a significant group, since most of them catalyze reactions essential for supporting the central metabolism, or are necessary for the pathogen vitality. Genomic exploration of pathogenic and non-pathogenic microorganisms has revealed genes encoding for a superfamily of metalloenzymes, known as carbonic anhydrases (CAs, EC 4.2.1.1). CAs catalyze the physiologically crucial reversible reaction of the carbon dioxide hydration to bicarbonate and protons. Herein, we investigated the sulfonamide inhibition profile of the recombinant β-CA (CynT2) identified in the genome of the Gram-negative bacterium Escherichia coli. This biocatalyst is indispensable for the growth of the microbe at atmospheric pCO2. Surprisingly, this enzyme has not been investigated for its inhibition with any class of CA inhibitors. Here, we show that CynT2 was strongly inhibited by some substituted benzene-sulfonamides and the clinically used inhibitor sulpiride (KIs in the range of 82–97 nM). This study may be relevant for identifying novel CA inhibitors, as well as for another essential part of the drug discovery pipeline, such as the structure–activity relationship for this class of enzyme inhibitors.

The gram-negative bacterium Escherichia coli as a model for testing the effect of carbonic anhydrase inhibition on bacterial growth

“…Glucose was added as the only carbon source to a liquid minimal culture media for bacterial growth. It has been demonstrated earlier by our groups that AZA was able to inhibit in vitro the two recombinant E. coli CAs, β-CA (CynT2) and γ-CA (EcoCAγ), with K I s of 227 and 248 nM, respectively 40 , 41 .…”

Selective Inhibition of Helicobacter pylori Carbonic Anhydrases by Carvacrol and Thymol Could Impair Biofilm Production and the Release of Outer Membrane Vesicles

“…This strategy has been applied to different Gram-positive and Gram-negative bacterial species ( Escherichia coli , H. pylori , Mycobacterium tuberculosis , Vibrio cholerae , Pseudomonas aeruginosa , Staphylococcus aureus , Porphyromonas gingivalis , Streptococcus spp., Enterococcus spp., etc. ), hampering their ability to develop resistance to common antimicrobials and leading to innovative drug combinations to overcome the spreading of resistant strains [ 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 ].…”

Novel 1,3,5-Triazinyl Aminobenzenesulfonamides Incorporating Aminoalcohol, Aminochalcone and Aminostilbene Structural Motifs as Potent Anti-VRE Agents, and Carbonic Anhydrases I, II, VII, IX, and XII Inhibitors

“…Inhibition of bacterial CAs leads to the inhibition of growth, growth defects, and makes bacteria vulnerable to host defense mechanisms [ 4 , 8 , 9 ]. Over the last few years, a number of articles describing the inhibition activity of clinically licensed or new sulfonamides against bacterial CAs presented in Vibrio cholerae [ 10 , 11 , 12 , 13 , 14 ], Mycobacterium tuberculosis [ 15 , 16 , 17 , 18 ], Burkholderia pseudomallei [ 19 ], Burkholderia territorii [ 7 , 20 ], Escherichia coli [ 21 , 22 ], and others, were published.…”