Green synthesis of nanoparticles using plant extracts has emerged as an eco-friendly and cost-effective alternative to conventional chemical synthesis methods. Aqueous extract of Musa paradisiaca L. inflorescence contains bioactive compounds such as polyphenols, flavonoids, alkaloids and tannins etc and acts as a natural reducing and stabilizing agent. In the present study, Aqueous extract of Musa paradisiaca L. cultivar puttabale inflorescence were subjected for nanoparticles synthesis, characterization through UV-Visible spectroscopy, Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM). Dynamic light scattering (DLS), particle size zeta potential and evaluate the antimicrobial activity. The results confirmed the formation of iron nanoparticles with nanoscale size and stable morphology. The antimicrobial activity of both aqueous extract and synthesised nanoparticles showed good zone of inhibition against bacteria namely Staphylococcus aureus & Pseudomonas aeruginosa and fungus namely Aspergillus flavus & Aspergillus Niger. The study demonstrates that banana inflorescence extract can be effectively utilized for green synthesis of iron nanoparticles, contributes the various biomedical applications and environmental remediation