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Investigation of the antibacterial potential of (R)-(+) limonene against food-associated strains of pseudomonas aeruginosa through in vitro and in silico analyses

Millena de Souza Alves, Maria Alice Araújo de Medeiros, Renata Kelly Silva Medeiros, Cássio Ilan Soares Medeiros, Layane Alves Ramalho, Gabrielle Nóbrega Alves, Wesley Kauã da Silva Lima, Maria Heloísa Furtunato Rocha et al.

Agrobiological Records, 2026

Abstract

Antimicrobial resistance is one of the major global health challenges.Consequently, the effectiveness of conventional antimicrobials has progressively declined due to the emergence of multidrug-resistant strains, making the search for antimicrobial alternatives essential.This study aimed to evaluate the antimicrobial activity of (R)-(+)-limonene against Pseudomonas aeruginosa strains isolated from food and to investigate its possible mechanism of action via molecular docking with DNA gyrase B. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays were performed using microdilution in 96-well plates, while the Minimum Inhibitory Concentration for Adhesion (MICA) was assessed in test tubes.In addition, interactions with antimicrobials were investigated using the disk diffusion method, and molecular docking was performed with AutoDock 4.2.The results showed an MIC90 of 1000 μg/mL and an MBC at ratios of 1:1 and 2:1, indicating a bactericidal effect.Regarding anti-adhesive activity, (R)-(+)-limonene inhibited biofilm formation at a dilution ratio of up to 1:8.With regard to combination with antimicrobials, a synergistic effect was observed with ampicillin, gentamicin, tetracycline, ciprofloxacin, ceftazidime, and cefazolin.Additionally, interactions with the active site of the DNA gyrase B enzyme were identified, primarily via hydrophobic, van der Waals, and alkyl/pi-alkyl interactions.These findings reinforce the potential of…

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