In Silico Analysis of Terpinen-4-ol as a Sustainable Anti-Biofilm Agent Targeting E. coli Outer Membrane Protein OmpX

Terpinen-4-ol, a significant component of Melaleuca alternifolia essential oil, was examined for its potential as an anti-biofilm agent against E. coli to enhance sustainable food preservation. Terpinen-4-ol demonstrated potent antibacterial activity against E. coli, with an MIC of 0.39%. Confocal microscopy revealed that concentration lower than the MIC successfully disrupted the 3D biofilm architecture, transforming dense matriced into distinct colonies. Swiss Dock molecular docking simulations revealed a strong binding interaction between terpinen-4-ol and the E. coli outer membrane protein OmpX, with the most stable configuration showing a binding affinity of -5.70 kcal/mol. Molecular dynamics simulations of the OmpX-terpinen-4-ol complex over 100 ns using GROMACS confirmed the stability of the protein-ligand system, as indicated by low RMSD values, a consistent radius of gyration, and intermittent hydrogen bonding. In silico ADME predictions via SwissADME indicated pharmacokinetic properties for terpinen-4-ol, including a molecular weight of 206.28 g/mol, a log P range of 2.5 to 3.4, and a TPSA of 37.30 Ų, suggesting good bioavailability and membrane permeability. Cytotoxicity predictions in human cell lines using CLC-Prd showed low toxicity probabilities. Hydrophobicity plot analysis of OmpX (PDB ID: 1QJ8) revealed a unique eight-stranded antiparallel β-barrel structure, with hydrophobic residues on the outer surface interacting with the lipid bilayer and hydrophilic residues lining the inner pore. These computational results underscore the potential of terpinen-4-ol as a promising anti-biofilm agent for sustainable food preservation, further experimental validation, and application in the food industry.