Pure and Fe-doped ZnO nanostructures with different weight ratios (0.5, 1.0, 1.5, and 2.0 at wt% of Fe) were successfully synthesized by a facile microwave combustion method using urea as a fuel. The detailed structural characterization was performed by means of X-ray diffraction (XRD), high resolution scanning electron microscopy (HR-SEM), energy dispersive X-ray analysis (EDX), diffuse reflectance spectroscopy (DRS), photoluminescence (PL) spectroscopy and vibrating sample magnetometry (VSM). XRD patterns refined by the Rietveld method indicated that Fe-doped ZnO have a single pure phase with wurtzite structure, suggesting that Fe ions are successfully incorporated into ZnO crystal lattice by occupying Zn ionic sites. Interestingly, the morphology was found to change substantially from grains to nanoflakes and then into nanorods with the variation of Fe-content. The optical band gap estimated using DRS was found to be red-shifted from 3.220 eV for the pure ZnO nanostructures, then decreases up to 3.200 eV with increasing Fe-content. Magnetic studies showed that Fe-doped ZnO nanostructures exhibit room temperature ferromagnetism (RTFM) and the saturation magnetization attained a maximum value of 8.154×10-3 emu/g for the highest Fe-content. The antibacterial activity of pure and Fe-doped ZnO nanostructures against a Gram-positive bacteria and Gram-negative bacteria was investigated. Pure ZnO and Fe-doped ZnO exhibited antibacterial activity, but it was considerably more effective in the 1.5 wt% Fe-doped ZnO nanostructures. ©2016 American Scientific Publishers All rights reserved.