Nanotechnology is the science, engineering and technology conducted at the scale that ranges between 1-100 nanometers. For the bio-application, evolution of nanotechnology is creating the concern of scientists towards the synthesis of nanoparticles. The nanoparticles have unique characteristics as compare to bulk materials. Zinc oxide (ZnO) is a matchless semiconductor and it has been under investigation due to its wide range of applications in various areas like biomedical, electronics, material science and optics. In the present work synthesis of ZnO nanoparticles was carried out by using simple chemical approach, Sol-gel method for being effective and inexpensive, by employing zinc acetate dehydrate Zn (CH3CO2)2.2H2O as a precursor and sodium hydroxide (NaOH) starch as a constant agent. The structural properties of resultant zinc oxide nanoparticles were investigated by X-ray diffraction (XRD) technique. The XRD data confirmed the hexagonal wurtzite structure of ZnO powder confirmed by JCPDS 36-1451 data. Particles size was calculated by Scherrer formula and calculated size was 30.14 nm. These nanoparticles were investigated for inhibition zone of bacterial strain Escherichia coli, a gram-negative microbe, at various concentrations of ZnO nanoparticles. Zinc oxide nanoparticles were very proficient for inhibition of growth of bacterial strain E. coli. The mechanism of ZnO NPs for antibacterial activity is release of reactive oxygen species which not only hydrolyze cell wall but cell membrane and cellular components as well providing a potential bactericidal effect.