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Research Article

In-silico and in-vitro assessment of the antibiofilm potential of azo dye, carmoisine against Pseudomonas aeruginosa

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Received 01 Mar 2023, Accepted 07 Jul 2023, Published online: 24 Jul 2023
 

Abstract

Biofilm is a community of microorganisms attached to the substrate and plays a significant role in microbial pathogenesis and medical device-related infection. Pseudomonas aeruginosa (PA) is a highly infectious gram-negative opportunistic biofilm-forming bacterium with high antibiotic resistance. Several reports underscore the antimicrobial activity of natural and synthetic food coloring agents, including carmoisine, turmeric dye, red amaranth dye, and phloxine B. However, their ability to suppress the PA biofilm is not clearly understood. Carmoisine is a red-colored synthetic azo dye containing naphthalene subunits and sulfonic groups and is widely used as a food coloring agent. This study investigated the antibiofilm potential and possible mechanism of biofilm inhibition by carmoisine against PA. Computational studies through molecular docking revealed that carmoisine strongly binds to QS regulator LasR (-12.7) and relatively less strongly but significantly with WspR (-6.9). Further analysis of the docked LasR-carmoisine complex using 100 ns MD simulation (Desmond, Schrödinger) validated the bonding strength and stability. Crystal violet assay, triphenyl tetrazolium chloride salt assay, and confocal microscopic studies were adopted for biofilm quantification, and the results indicated the dose-dependent antibiofilm activity of carmoisine against PA. We hypothesise that the carmoisine-mediated reduction of biofilm in PA is due to its interaction with LasR and interference with the QS system. The computational and biochemical analysis of another compound, 1,2-naphthoquinone-4-sulphonic acid, reiterated the role of the naphthalene ring in biofilm inhibition. Hence, this work will pave the way for the future discovery of antibiofilm drugs based on naphthalene ring-based lead compounds.

Communicated by Ramaswamy H. Sarma

Acknowledgments

The authors are extremely grateful to Dr Sajeeb Khan, Department of Zoology, Government College for Women, Thiruvananthapuram, Kerala (India), for providing the necessary facilities. The University Grant Commission, Junior Research Fellowships, awarded to Minsa Mini and Devi Jayakumar, is duly acknowledged. We also acknowledge the contribution of Karthika Suryalatha and Sabu Thomas, RGCB, for microscopy works.

Disclosure statement

No potential conflict of interest was reported by the authors.

Author contribution

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Minsa Mini and Praveen Kumar. The first draft of the manuscript was written by Minsa Mini, and Devi Jayakumar and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Additional information

Funding

Research funding under the Performance Linked Encouragement for Academic Studies and Endeavour (P LEASE) Scheme, Kerala Government and Consolidation of University Research for Innovation and Excellence in Women Universities (CURIE), DST, New Delhi.

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