In the present work, Zn1-xAgxO (x = 0, 0.03, 0.05, and 0.07) nanoparticles were synthesized by the sol-gel synthesis method. Microstructural, morphological, optical, and antibacterial characterizations were performed. The wurtzite crystalline structure was confirmed through XRD analysis. The maximum crystallite size was found for Zn0.93Ag0.07O (∼22.36 nm). Rietveld refinement was performed to understand better crystal symmetry. The optical band gap property exhibited a decremental pattern with the increasing Ag content. The lowest band gap was achieved by Zn0.93Ag0.07O (∼3.26 eV). The Fourier transform infrared spectroscopy (FTIR) analysis ensured the bond stretching of Zn–O and Ag–O near the absorbance band of 490 cm−1. Further, the Thermogravimetric analysis (TGA) was performed to determine the samples' weight loss, and the minimum weight loss was observed for Zn0.93Ag0.07O (∼9–11%). Finally, the antibacterial characterization was performed using Escherichia coli (gram-negative bacteria), and again Zn0.93Ag0.07O exhibited better performances than other synthesized nanoparticles.