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The current study demonstrates a sophisticated and environmentally friendly synthesis of zinc oxide nanoparticles (ZnO-NPs) for a range of biological and environmental applications using <i>Monotheca buxifolia</i> as a bio-source. At the nanometer scale, a simple aqueous extract from <i>Monotheca buxifolia</i> was used to convert Zn into stable elemental zinc (Zn⁰). With an average size of 45.8 nm and a spherical shape, the NPs were stable and pure. The nanoparticles studied here were tested in vitro for bactericide, fungicide, biocompatibility, leishmaniasis, anti-diabetic effect, antioxidant effect, and anti-Alzheimer’s effect. According to our results, <i>Monotheca buxifolia</i> mediated ZnO-NPs are highly effective against spore-forming fungal strains and MDR bacterial strains. All examined bacterial isolates of UTI (urinary tract infection) were resistant to non-coated antibiotics; however, adding 1% of the produced ZnO-NPs to the treatments increased their bactericidal activity significantly. The NPs also showed dose-dependent cytotoxicity against Leishmania tropica parasites, with an LC50 of 248 μg/mL for promastigote parasites and 251 μg/mL for amastigote parasites. In addition, a significant inhibition of α-glucosidase, α-amylase, butyrylcholinesterase (BChE), and acetylcholinesterase (AChE) was discovered, indicating anti-Alzheimer’s and anti-diabetic effects. The biocompatibility of the particles with human red blood cells was also observed. Due to their environmentally friendly production, biological safety, and exceptional physicochemical properties, ZnO-NPs could be used as a new competitor for several biological and environmental applications.