The perpetual use of antibiotics against pathogens inadvertently altered their genes that have translated into an unprecedented resistance in microorganisms in the twenty-first century. Many researchers have formulated bactericidal and bacteriostatic inorganic nanoparticle-based antiseptics that may be linked to broad-spectrum activity and far lower propensity to induce microbial resistance than organic-based antibiotics. Based on this line, herein, we present observations on microbial abatement using gold-based zinc oxide nanostructures (Au@ZnO) which are synthesized using hydrothermal route. Inhibition of microbial growth and biofilm using Au@ZnO is a unique feature of our study. Furthermore, this study evinces antimicrobial and antibiofilm mechanisms of photo-eradiated Au@ZnO by disruption of cellular functions and biofilms via reactive oxygen species (ROS)-dependent generation of superoxide anion radical. The present study is significant as it introduces novel functionalities to Au@ZnO in the biomedical field which can be extended to other species of microbial pathogens.