Green metal nanoparticles (NPs) have emerged as efficient biofilm inhibitors. Citrus macroptera (CM) fruit extract mediated iron (Fe) NPs, i. e., CM-FeNPs were thus synthesized for studying biofilm inhibition property. The shape (spherical), size (12 nm), and crystallinity (α-Fe 0 bcc) of the CM-FeNPs were confirmed by studying mainly Transmission Electron microscopy (TEM) and X-ray Diffractometer (X-RD) tools. The particles inhibit biofilm growth of a bacteria Pseudomonas aeruginosa (P. aeruginosa) by inhibiting the functions of extracellular polymeric substances (EPS) and virulence factors to ∼ 80 %. Scanning Electron microscopy (SEM) images of the bacteria in presence of the CM-FeNPs (4600 nM) in combination with azithromycin (AZI) evidence the inhibition of biofilm growth > 80 %. Molecular docking studies support experimental findings and result significant negative binding energies (e. g., isoplatydesmine/LasR: À 8.25 kcal/mole, isoplatydesmine/MvfR: À 7.56 kcal/mole) from bioactive molecules/biofilm proteins interactions. Thus, CM-FeNPs could be considered as potential biofilm inhibitors, and as an alternative to antibiotic drugs.