CeO2 nanoparticles (NPs) were green synthesized using Gloriosa superba L. leaf extract. The synthesized nanoparticles retained the cubic structure, which was confirmed by X-ray diffraction studies. The oxidation states of the elements (C (1s), O (1s) and Ce (3d)) were confirmed by XPS studies. TEM images showed that the NPs possessed spherical shape and particle size of 5 nm. The Ce–O stretching bands were observed at 451 cm− 1 and 457 cm− 1 from the FT-IR and Raman spectra respectively. The band gap of the CeO2 NPs was estimated as 3.78 eV from the UV–visible spectrum. From the photoluminescence measurements, the broad emission composed of eight different bands were found. The antibacterial studies performed against a set of bacterial strains showed that Gram positive (G +) bacteria were relatively more susceptible to the NPs than Gram negative (G −) bacteria. The toxicological behavior of CeO2 NPs was found due to the synthesized NPs with uneven ridges and oxygen defects in CeO2 NPs.