This study presents a green synthesis approach for the production of copper oxide nanoparticles (CuONPs) using Plumbago auriculata Lam. leaf extract. The synthesized CuONPs were characterized using various techniques, including UV-Vis spectrometry, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM). The physicochemical properties of the CuONPs were thoroughly analyzed, revealing their unique biological and mechanical properties. Furthermore, the antimicrobial efficacy of the CuONPs was evaluated against several pathogenic bacterial strains and Candida albicans using the disc diffusion method and determination of minimum inhibitory concentration (MIC). The CuONPs exhibited significant antimicrobial activity, highlighting their potential as antimicrobial agents. Additionally, the antioxidant activity of the CuONPs was assessed through DPPH scavenging activity testing, indicating their ability to counteract free radicals and potential application in managing oxidative stress-related disorders. Overall, this study contributes to the development of ecofriendly and biocompatible CuONPs with potential applications in medicine, agriculture, and environmental remediation