Nanostructured particles offer outstanding diversities of applications in the fields of nanotechnology, nano-engineering, nano-biotechnology, etc. Morphological structure, size distribution, electronic behavior including intrinsic characteristics of nanoparticles depend on the source and synthesis methods. Here, an eco-friendly approach using microwave irradiation for the synthesis of zinc oxide (ZnO) nanoparticles has been reported. Zinc nitrate was used as a precursor whereas starch and D-glucose were used as capping and reducing agent, respectively. The synthesized nanoparticles were characterized by different instrumental methods including Ultraviolet-Visible spectroscopy (UV-Vis), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDX), and field emission scanning electron microscopy (FE-SEM). The characteristic λ at 373 nm for ZnO nanoparticles was recorded from UV-Vis absorption spectrum in aqueous system. FT-IR spectrum showed a very sharp peak at 476.62 cm which confirmed the presence of Zn-O bond. The prepared ZnO was highly crystalline having wurtzite structure and the crystallite size was calculated to be 24.41 nm obtained from XRD analysis. FE-SEM images showed that the synthesized ZnO nanoparticles had near- spherical morphology and particle size was found in the range of 40-90 nm. The antibacterial and anti-biofilm application of ZnO nanoparticles were studied and inhibition zones of Gram negative Salmonella typhi (S. typhi), Klebsiella spp., Escherichia coli (E. coli) and Gram positive Staphylococcus aureus (S. aureus)- 8a were found to be 11 mm, 12 mm, 11.5 mm and 13.5 mm, respectively. Besides, ZnO nanoparticles also showed excellent photocatalytic activity against methylene blue dye solution. The easy and eco-friendly fabrication method would play vital role in other nanoparticles synthesis to meet the demand in textile industry, agriculture and medical sectors.