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Deciphering the antibiofilm potential of 2-Phenylethyl methyl ether (PEME), a bioactive compound of Kewda essential
Priya Cheruvanachari1, Subhaswaraj Pattnaik1, Monika Mishra1
1Centre of Excellence in Natural Products and Therapeutics, Department of Biotechnology and Bioinformatics, Sambalpur University, Jyoti Vihar, Sambalpur, 768 019, Odisha, India.
Abstract:
Opportunistic pathogenic bacteria and their pathogenicity linked with biofilm infections become a severe issue as they resist the actions of multiple antimicrobial drugs. Naturally derived drugs having antibiofilm properties are more effective than chemically synthesized drugs. The plant derived essential oils are a rich source of phytoconstituents with widespread pharmacological values. In the present investigation, a major phytoconstituent, 2-Phenyl Ethyl Methyl Ether (PEME) of Kewda essential oil extracted from the flowers of Pandanus odorifer was explored for its prospective antimicrobial and anti-biofilm properties against ESKAPE pathogenic bacterial strains, Staphylococcus aureus and MTCC 740. The minimum inhibitory concentration (MIC) of PEME was found to be 50 mM against the tested bacterial strains. A gradual decrease in biofilm production was observed when PEME was treated with the sub-MIC concentration. The reduction in biofilm formation was noticeable from qualitative assay i.e., Congo Red Agar Assay (CRA) and further quantified by crystal violet staining assay. The decline in exopolysaccharides production was quantified, with the highest inhibition against MTCC 740 with a decrease of 71.76 ± 4.56% compared to untreated control. From the microscopic analysis (light and fluorescence microscopic method), PEME exhibited inhibitory effect on biofilm formation on the polystyrene surface. The in silico studies stated that PEME could invariably bind to biofilm associated target proteins. Further, transcriptomic data analysis suggested the role of PEME in the down-regulation of specific genes, agrA, sarA, norA and mepR, which are critically associated with bacterial virulence, biofilm dynamics and drug resistance patterns in S. aureus. Further, qRT-PCR analysis validated the role of PEME on biofilm inhibition by relative downregulation of agrA, sarA, norA and mepR genes. Further, advanced in silico methodologies could be employed in future investigations to validate its candidature as promising anti-biofilm agent.
Insights
2-Phenyl Ethyl Methyl Ether (PEME), a compound from Kewda essential oil, effectively inhibits pathogenic bacterial biofilm formation. This natural compound shows promise as an antimicrobial and anti-biofilm agent against drug-resistant bacteria.
Area of Science:
- Microbiology
- Pharmacology
- Natural Products Chemistry
Background:
- Opportunistic bacterial pathogens form biofilms, leading to severe infections resistant to multiple antimicrobial drugs.
- Naturally derived compounds, particularly plant-derived essential oils, offer effective antibiofilm properties compared to synthetic drugs.
- Kewda essential oil from Pandanus odorifer flowers contains valuable phytoconstituents with pharmacological potential.
Purpose of the Study:
- To investigate the antimicrobial and anti-biofilm potential of 2-Phenyl Ethyl Methyl Ether (PEME), a major phytoconstituent of Kewda essential oil.
- To evaluate PEME's efficacy against ESKAPE pathogenic bacterial strains, including Staphylococcus aureus.
- To elucidate the molecular mechanisms underlying PEME's anti-biofilm activity.
Main Methods:
- Minimum Inhibitory Concentration (MIC) determination for PEME against bacterial strains.
- Qualitative (Congo Red Agar Assay) and quantitative (crystal violet staining) assessment of biofilm inhibition.
- Microscopic analysis (light and fluorescence microscopy) of biofilm formation on polystyrene surfaces.
- In silico molecular docking studies to predict PEME's binding to biofilm-associated proteins.
- Transcriptomic analysis and qRT-PCR to identify and validate PEME's effect on key bacterial genes (agrA, sarA, norA, mepR).
Main Results:
- PEME demonstrated antimicrobial activity with an MIC of 50 mM against tested strains.
- Sub-MIC concentrations of PEME significantly reduced biofilm production, with notable inhibition of exopolysaccharides (up to 71.76% against MTCC 740).
- Microscopic analyses confirmed PEME's inhibitory effect on biofilm formation.
- In silico studies suggested PEME binds to biofilm-associated target proteins.
- Transcriptomic and qRT-PCR analyses revealed PEME down-regulates critical genes (agrA, sarA, norA, mepR) involved in virulence, biofilm formation, and drug resistance in S. aureus.
Conclusions:
- 2-Phenyl Ethyl Methyl Ether (PEME) exhibits significant antimicrobial and anti-biofilm properties against pathogenic bacteria.
- PEME effectively inhibits biofilm formation by down-regulating key virulence and resistance genes.
- PEME represents a promising natural candidate for developing novel anti-biofilm therapeutic strategies.
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