Gold nanoparticles (AuNPs) have been known to possess exceptional electric, biochemical, and optical characteristics and are ‘the topic of discussion’ these days, especially relating to the field of biomedicine. Several plants, bacteria, and fungi have been utilized for the generation of AuNPs, besides other physical and chemical methods. While some studies have been reported with gold nanoparticles, less are aimed at fungi and its optimization factors. These parameters can allow us to design AuNPs of our choice depending on the use. The present review focuses on and inspects AuNPs with green synthesis through fungus optimization parameters followed by applications, aiming specifically at their antibacterial activity. Their antibacterial characteristics can open new doors for the pharmaceutical industry in the future.