In the present study Mentha arvensis medaited Magnesium oxide nanoparticles were synthesized by novel green route followed by advanced characterization via XRD, FTIR, UV, SEM, TEM, DLS and TGA. The mean grain size of 32.4 nm and crystallite fcc morphology were confirmed by X-ray diffractive analysis. Scanning and Transmission electron microscopy analysis revealed the spherical and elliptical morphologies of the biosynthesized nanoparticles. Particle surface charge of −16.1 mV were determined by zeta potential and zeta size of 30–120 nm via dynamic light scattering method. Fourier transform spectroscopic analysis revealed the possible involvement of functional groups in the plant extract in reduction of Mg2+ ions to Mg0. Furthermore, the antioxidant, anti-Alzheimer, anti-cancer, and anti-H. pylori activities were performed. The results revealed that MgO-NPs has significant anti-H. pyloric potential by giving ZOI of 17.19 ± 0.83 mm against Helicobacter felis followed by Helicobacter suis. MgO-NPs inhibited protein kinase enzyme up to 12.44 ± 0.72% at 5 mg/mL and thus showed eminent anticancer activity. Significant free radicals scavenging and hemocompatability was also shown by MgO-NPs. MgO-NPs also displayed good inhibition potential against Hela cell lines with maximum inhibition of 49.49 ± 1.18 at 400 µg/mL. Owing to ecofriendly synthesis, non-toxic and biocompatible nature, Mentha arvensis synthesized MgO-NPs can be used as potent antimicrobial agent in therapeutic applications.