Pullulan (Pull) decorated with monodisperse Ag and Au nanoparticles (NPs) was synthesized by a simple and green method. Samples were characterized by FTIR, UV-vis, NMR, XRD, TGA, SEM, XPS, DLS, and TEM. SEM images showed highly oriented microforms reported for the first time for Pull, because of the supramolecular self-assembling behavior of Pull chains. Antimicrobial and quorum sensing (QS) inhibition activities were tested against six pathogen bacteria and reporter and biomonitor strain. Pull decorated with NPs, in particular, Ag-modified ones, outperformed pristine Pull. The cell proliferation was tested with an MTT assay. NPs-decorated Pull was studied for the first time as an inhibitory agent against bacterial signal molecules and found to be a good candidate. The promising performance of AgNPs@Pull compared to the commercial antibiotic gentamicin showed that it has great potential as a therapeutic approach to overcome the bacterial resistance that has developed against conventional antibiotics.