Green synthesis and antibacterial-antibiofilm properties of biogenic silver nanoparticles

The biosynthesis of metallic nanoparticles is gaining prominence as an alternative to traditional physicochemical methods, offering several advantages such as simplicity, non-toxicity, lower energy requirements and short reaction times leading to environmentally sustainable processes. The aims of this work were: to study the extracellular biosynthesis of silver nanoparticles (AgNPs) by Pseudomonas extremaustralis 2E-UNGS, to characterise the shape, monodispersity and size of AgNPs, to explore their antimicrobial and antibiofilm activities, and to evaluate the role of nitrate reductase activity in the biosynthesis process. The novelty of this work relies on the development of a green and sustainable method for the synthesis of stable AgNPs with optimal properties for potential applications in antimicrobial materials, especially when incorporated into polymeric matrices or used as agrochemical substitutes. Optimal conditions for the biosynthesis of spherical AgNPs were determined to be pH 7, 38 °C, 4 h of darkness and 120 rpm using stationary phase culture supernatants of P. extremaustralis 2E-UNGS. The involvement of extracellular nitrate reductase in AgNP biosynthesis was confirmed by enzymatic assays and supported by bioinformatics analysis, which identified the presence of the napA2 gene linked to the nirBD cluster. Antimicrobial assays demonstrated the inhibitory effect of AgNPs against both Gram-positive and Gram-negative bacteria, including Pseudomonas aeruginosa PA01 in both planktonic and biofilm states. In addition, the potential application of AgNPs in innovative antibacterial polymers was explored by incorporating them into polyurethane matrices either alone (PU-AgNP) or in combination with crystal violet as a photosensitizer (PU-AgNP-CV). Subsequent inoculation with a clinical isolate of Pseudomonas aeruginosa resulted in significant reductions in viable bacterial counts on both PU-AgNP-CV and PU-AgNP. Biogenic AgNPs showed antibacterial and antibiofilm properties for new antimicrobial material development.