In this study, extracellular biosynthesis of zinc oxide nanoparticles (ZnO NPs) by using the supernatant of Bacillus subtilis ZBP4 after cultivation in nutrient broth at 33 °C for 24 h was investigated. Zinc sulfate heptahydrate was used as the precursor, and the reactions were performed at 33 °C for 72 h. The effects of pH 5-9 and precursor concentration (2-10 mM) were determined and the optima were found to be pH 7.5 and 8 mM ZnSO4·7H2O, respectively. The nanoparticles were characterized by UV-VIS spectroscopy, FESEM, TEM, EDS, XRD, FTIR and zeta potential measurement. ZnO NPs appeared to be irregular spherical structures with a size range of 22-59 nm, as confirmed by FESEM and TEM. Energy-dispersive spectroscopy analysis validated the formation of ZnO NPs. X-ray diffraction analysis revealed the crystalline structure of the ZnO NPs, and they were determined to have a zeta potential of -19.0 ± 4.3 mV and a bandgap of 3.36 eV. Antibacterial activity experiments showed that ZnO NPs are effective against a broad spectrum of Gram-positive and Gram-negative food pathogens. This study provides evidence for a safe and effective method for synthesizing ZnO NPs and demonstrates their effectiveness against pathogenic bacteria.
Keywords: Antibacterial activity; Bacillus; ZnO NPs; biosynthesis; nanoparticles.