Vanadium oxide (V 2 O 5 ) and carbon spheres (Cs)-doped NiO 2 nanostructures (NSs) were prepared using the co-precipitation approach. Several spectroscopic and microscopic techniques, including X-ray diffraction (XRD), UV−vis, FTIR, TEM, and HR-TEM investigations, were used to describe the as-synthesized NSs. The XRD pattern exhibited the hexagonal structure, and the crystallite size of pristine and doped NSs was calculated as 29.3, 32.8, 25.79, and 45.19 nm, respectively. The control sample (NiO 2 ) showed maximum absorption at 330 nm, and upon doping, a redshift was observed, leading to decreased band gap energy from 3.75 to 3.59 eV. TEM of NiO 2 shows agglomerated nonuniform nanorods exhibited with various nanoparticles without a specific orientation; a higher agglomeration was observed upon doping. The (4 wt %) V 2 O 5 /Cs-doped NiO 2 NSs served as superior catalysts with a 94.21% MB reduction in acidic media. The significant antibacterial efficacy was estimated against Escherichia coli by measuring the zone of inhibition (3.75 mm). Besides their bactericidal analysis, V 2 O 5 /Cs-doped NiO 2 was shown to have a binding score of 6.37 for dihydrofolate reductase and a binding score of 4.31 for dihydropteroate synthase in an in silico docking study of E. coli.