Multi-faceted exploration of the novel active γ-carbonic anhydrase PaCAγ1 in the human pathogen Pseudomonas
Vincenzo Massimiliano Vivenzio1, Reygan Braga2, Alessandro Bonardi3
1Istituto di Biostrutture e Bioimmagini, CNR, Via Pietro Castellino 111, 80131 Napoli, Italy; Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania "Luigi Vanvitelli", 81100 Caserta, Italy.
Abstract:
Pseudomonas aeruginosa, a ubiquitous Gram-negative bacterium, is an opportunistic pathogen that causes severe health issues in both immunocompetent and immunocompromised individuals. It is a member of the ESKAPE group of pathogens, and a leading cause of nosocomial infections worldwide. With the aim of identifying novel tools to combat P. aeruginosa infections, and considering that carbonic anhydrases are emerging as promising targets to overcome antibiotic resistance, we present here the identification and detailed characterization of a γ-class carbonic anhydrase, PaCAγ1, from P. aeruginosa using a multidisciplinary approach. Specifically, the identified enzyme was recombinantly produced and subjected to an extensive biochemical and biophysical characterization, proving its enzymatic function and revealing its overall stability at high temperatures. We determined that PaCAγ1 plays a role in pathogen fitness under carbon-limiting conditions and contributes to its survival within the macrophages. Various sulfonamide, sulfamide, and sulfamate derivatives were evaluated for their inhibitory properties against PaCAγ1 following a drug repurposing approach. This study reports on the discovery and characterization of a previously unrecognized γ-class CA, which enhances our understanding of the biochemical capacity and physiological importance of these enzymes in human pathogens.
Insights
Researchers identified a novel gamma-class carbonic anhydrase (PaCAγ1) in Pseudomonas aeruginosa. This enzyme is crucial for pathogen survival under nutrient-poor conditions and offers a potential new target for combating antibiotic resistance.
Area of Science:
- Microbiology
- Biochemistry
- Enzymology
Background:
- Pseudomonas aeruginosa is a major Gram-negative opportunistic pathogen causing significant nosocomial infections.
- It belongs to the ESKAPE group, known for its antibiotic resistance.
- Carbonic anhydrases are emerging as promising therapeutic targets against resistant bacteria.
Purpose of the Study:
- To identify and characterize a novel gamma-class carbonic anhydrase (PaCAγ1) from P. aeruginosa.
- To explore PaCAγ1 as a potential target for developing new anti-Pseudomonas aeruginosa strategies.
- To investigate the role of PaCAγ1 in bacterial fitness and survival.
Main Methods:
- Multidisciplinary approach including recombinant protein production.
- Extensive biochemical and biophysical characterization of the enzyme.
- Evaluation of various sulfonamide, sulfamide, and sulfamate derivatives as potential inhibitors.
Main Results:
- Discovery and detailed characterization of a previously unrecognized gamma-class carbonic anhydrase, PaCAγ1.
- Demonstration of PaCAγ1's enzymatic function and high-temperature stability.
- Evidence that PaCAγ1 is essential for P. aeruginosa fitness under carbon limitation and survival within macrophages.
Conclusions:
- PaCAγ1 is a significant enzyme for P. aeruginosa physiology and virulence.
- This study expands the understanding of carbonic anhydrases in human pathogens.
- PaCAγ1 represents a novel potential target for antimicrobial drug development against P. aeruginosa infections.
More Related Videos
11:35Replication of the Ordered, Nonredundant Library of Pseudomonas aeruginosa strain PA14 Transposon Insertion Mutants
Published on: May 4, 2018
08:34Generation of In-Frame Gene Deletion Mutants in Pseudomonas aeruginosa and Testing for Virulence Attenuation in a Simple Mouse Model of Infection
Published on: January 8, 2020
