Abstract
Quorum sensing (QS) is pivotal in coordinating virulence factors and biofilm formation in various pathogenic bacteria, making it a prime target for disrupting bacterial communication. Pseudomonas aeruginosa is a member of the “ESKAPE” group of bacterial pathogens known for their association with antimicrobial resistance and biofilm formation. The current antibiotic arsenal falls short of addressing biofilm-related infections effectively, highlighting the urgent need for novel therapeutic agents. In this study, we explored the anti-QS and anti-biofilm properties of theophylline against two significant pathogens, Chromobacterium violaceum and P. aeruginosa. The production of violacein, pyocyanin, rhamnolipid, and protease was carried out, along with the evaluation of biofilm formation through methods including crystal violet staining, triphenyl tetrazolium chloride assay, and fluorescence microscopy. Furthermore, computational analyses were conducted to predict the targets of theophylline in the QS pathways of P. aeruginosa and C. violaceum. Our study demonstrated that theophylline effectively inhibits QS activity and biofilm formation in C. violaceum and P. aeruginosa. In P. aeruginosa, theophylline inhibited the production of key virulence factors, including pyocyanin, rhamnolipid, protease, and biofilm formation. The computational analyses suggest that theophylline exhibits robust binding affinity to CviR in C. violaceum and RhlR in P. aeruginosa, key participants in the QS-mediated biofilm pathways. Furthermore, theophylline also displays promising interactions with LasR and QscR in P. aeruginosa. Our study highlights theophylline as a versatile anti-QS agent and offers a promising avenue for future research to develop novel therapeutic strategies against biofilm-associated infections.
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Acknowledgements
We acknowledge the research funding provided by the Performance Linked Encouragement for Academic Studies and Endeavour (Grant No: P3/24770/2020/DCE) scheme of the Kerala Government and the Consolidation of University Research for Innovation and Excellence in Women Universities (Grant No: DST/CURIE-PG/2022/50) program of DST, New Delhi. We also acknowledge the University Grant Commission for awarding Junior Research Fellowships to Minsa Mini, Devi Jayakumar, Parvathi Vaikkathillam and Kerala University Fellowship to Aparna Mohan. We thank Dr. Sabu Thomas from the Rajiv Gandhi Centre for Biotechnology for generously providing C. violaceum (Vps1) for this study. The centralized Instrumentation facility, Government College for Women, is duly acknowledged.
Funding
Performance Linked Encouragement for Academic Studies and Endeavour (Grant No: P3/24770/2020/DCE) scheme of the Kerala Government and the Consolidation of University Research for Innovation and Excellence in Women Universities (Grant No: DST/CURIE-PG/2022/50) program of DST, New Delhi.
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AM and PK conceived and planned the experiments. AM, PPR, DJ, MM, PV, and SA carried out the experiments. AM, PPR, PK, and MM planned and carried out the docking and simulations. All authors contributed to the interpretation of the results. All authors took the lead in writing the manuscript. All authors provided critical feedback and helped shape the research, analysis, and manuscript.
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Mohan, A., Rajan, P.P., Kumar, P. et al. Theophylline as a quorum sensing and biofilm inhibitor in Pseudomonas aeruginosa and Chromobacterium violaceum. Int Microbiol (2024). https://doi.org/10.1007/s10123-024-00487-w
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DOI: https://doi.org/10.1007/s10123-024-00487-w