The present study focused on accomplishing this phenomenon by employing Coleus forskohlii root to produce zinc oxide nanoparticles (ZnO NPs), characterizing those NPs using interpretive approaches, and elucidating high-priority antimicrobial and anti-cancer abilities. UV-Vis spectroscopy was utilized to inspect the synthesized particles, which exhibited a color change pattern throughout the synthesis and proved their broad peak at 375 nm. The capping and stability of nanoparticles were predominantly due to polyphenols, flavonoids, and primary amines. The PSA and Zeta potential investigations confirmed a small particle size with improved stability. XRD measurements indicated the crystalline nature of particles with an average length of 32.54 nm. According to the SEM data, the average size of the ZnO NPs was 33.08 nm, and their formation was spherical to hexagonal in shape. The HR-TEM investigation pointed out that the ZnO nanoparticles showcased a hexagonal shape with an average mean size of 31.54 nm. The element ratios, which stated 73.08% zinc and 26.92% oxygen, were substantiated by the EDX analysis. The biosynthesized ZnO NPs show remarkable antibacterial activity against B. subtilis, S. aureus, and E. coli. Strong antifungal activity against C. albicans, A. flavus, and A. fumigatus is shown by ZnO NPs. The evaluation of ZnO NPs' anticancer properties broadens the scope of this study. With an IC50 value of 36.16 mu g/ml, this experiment confirmed feasible anticancer activity against the human ovarian cancer cell line SKOV-3. The novel green synthesis approach defined here presents a cost-efficient alternative for nanoparticle production and also creates new avenues for their use in several biological fields that include cancer treatment, antimicrobial pharmacology, and targeted drug delivery.