This paper presents a fast and cost-effective synthesis method for obtaining nano scaled multifunctional dysprosium iron oxide. The effects of reaction time and stirring rate on the compound's chemical and physical properties are investigated as synthesis parameters. The structural, morphological, thermal, magnetic, and optical characterizations of the compound are extensively investigated by current universal instruments. It was found in an orthorhombic system as DyFeO3 (ICSD = 98-028-0091). Their particle sizes were calculated as 7–10 nm by transmitting electron microscopy. Vibration sample magnetometer was used to obtain M-H curves that showed superparamagnetic properties in low magnetic fields. The band gap value was calculated 3.6 eV for the sample preparing at 30 min by McClean analysis using reflectance graphics. The fluorescence spectroscopy indicated a sharp transition at 4F9/2→ 6H13/2 which confirms the Dy3+ ion acting as luminescence center. Additionally, the nanoparticles exhibited a remarkably high microbial activity against both gram-positive and gram-negative bacteria. These comprehensive characterizations contribute to a better understanding of the synthesized nanomaterials and its use of their biomedical potential applications.