Background: The ancient plant species ginkgo biloba is thought to provide numerous health benefits to all living creatures. This plant is very bioactive and has a wide range of chemical constituents. There are several proven pharmacological and therapeutic benefits of G. biloba. Objective: The objectives of the present study were to produce iron nanoparticles (FeNPs), isolate a natural compound from Ginkgo biloba leaves, and evaluate the antibacterial properties of the resultant nanoparticles (NPs). Materials and Methods: Five extracts of Ginkgo biloba leaves were produced through various methods: cold water extraction, hot water extraction, cold alcoholic extraction, hot alcoholic extraction, and 70% alcoholic extraction. FeNPs biosynthesis occurred by reacting these Ginkgo biloba leaf extracts with iron sulfate Fe2 (SO4)3. Analysis techniques employed included GC/MS, SEM, XRD, and UV-Vis analysis. Results: The iron nanoparticles (FeNPs) utilized in this using scanning electronic microscopy measurement and UV-VIS spectroscopy, which has a characteristic peak at 217nm, it was possible to detect irregular structures with dimensions of no more than 41 nm. AFM was also used in this study to examine the NPs-Fe's size and morphology. The FTIR measurement also revealed the principal iron oxide bands at 685 and 596cm-1, which are caused by Fe-O vibrational stretching. Bioactivity was also carried out against certain bacteria. Conclusions: Antimicrobial activity showed an inhibiting effect on a variety of harmful bacteria. Klebsiella spp., Staphylococcus epidermidis and Candida albicans.