Antimicrobial and Anti-Biofilm Activities of Medicinal Plant-Derived Honey Against ESKAPE Pathogens: Insights into β-Lactamase Inhibition via Metabolomics and Molecular Modeling Studies

The emergence of multidrug-resistant bacterial infections is a major global public health concern. Human health is in danger from microorganisms that have developed resistance to currently used drugs. Honey is well known for its significant activity against antibiotic-resistant bacteria. In this study, the antibacterial properties of honey from various botanical sources in Saudi Arabia against seven significant nosocomial and foodborne pathogens were investigated. The physicochemical properties of four Saudi honey samples—aloe honey (HO1) (Aloe vera L.), anise honey (HO2) (Pimpinella anisum L.), moringa honey (HO4) (Moringa oleifera Lam.), and acacia honey (HO5) (Acacia sp.)—were examined. In addition, they were screened for antibacterial activity against ESKAPE pathogens (Enterobacter faecalis, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella Typhimurium, Escherichia coli, and Enterobacter sp.) and anti-biofilm activity against four pathogenic bacteria strains: S. aureus, P. aeruginosa, S. typhimurium, and E. coli. 1H NMR profiling and multivariate analysis (PCA and PLS-DA) were performed. Aloe honey (HO1) was the most distinct sample based on MVDA and its antibacterial activity, and it exhibited anti-biofilm activity against most biofilm-forming microorganisms. Its metabolic profile was deduced using LC-MS, and the resulting annotated compounds were docked against several β-lactamase enzyme classes. The results reveal the potential of honey-derived compounds to inhibit β-lactamases due to the presence of gallic acid hexoside and rosmarinic acid, suggesting their potential as competitive inhibitors. Our findings suggest that further honey antibacterial compounds could offer a novel approach to overcoming antibiotic resistance by targeting and inhibiting β-lactamase enzymes.