Nanotechnology has gained a successful stature as one of vital research areas in the twenty-first century. It has been proven that fixed nanomaterials has a high affinity for interaction with biological macromolecules through bonding to macromolecules, leading to lethal inactivation of bacteria and viruses. This study attempted to examine the effect of zinc oxide nanoparticles on the growth of E. coli bacteria in carrot juice. In fact, it focused on how zinc oxide nanoparticles (ZnO) at concentrations of 1 mM, 3mM, 5mM and 8mM affect the growth of E. coli bacteria in agar medium. Then, a spectrophotometer was used to measure the wavelength of E. coli at concentrations of 3mM, 1mM, 5mM and 8mM followed by dilution. The results showed that E. coli had the lowest absorption coefficient at 5 and 8 mM. Dilution of carrot juice revealed that the number of colonies decreased over time as the lowest number of colonies was observed at concentration of 8 mM within 48 hour. Given the antibacterial properties of zinc oxide nanoparticles, it is a modern technique in the food industry to apply nanoparticles to combat pathogenic microbial agents. It is expected with the application of zinc oxide nanoparticles at allowable doses will be common practice within a few years.