Zinc oxide (ZnO) is a wide band gap semiconductor (3.2eV) with a high exciton binding energy (60meV), where it has wide applications in advanced spintronic devices. The theoretical prediction of room temperature ferromagnetism and also antibacterial activity will be possible through the investigation of diluted magnetic semiconductors (DMS), such as transition metal doped ZnO, especially Cobalt doped ZnO. The aim of the work is the synthesis of Cobalt (Co) doped ZnO nanopowders were prepared Zn1−xCoxO (0⩽x⩾0.09) nanopowders from Sol–Gel auto combustion method have been synthesized with precursors such as Zinc and Cobalt nitrates with the assistance Ammonium acetate & Urea as fuel by increasing the cobalt concentration in zinc oxide and their structural, morphological, optical, Thermal, magnetic and antibacterial properties were studied by X-ray diffraction (XRD), field emission scanning electron microscope (FESEM), Transmission Electron microscope (TEM), UV–visible spectroscopy, thermo gravimetric/differential thermal analysis (TG/DTA) and vibrating sample magneto meter (VSM). From the antibacterial studies, against gram positive Bacillus subtilis bacteria is most abundant bacteria in soil and indoor atmosphere, which affects the stored spintronic devices so that the devices should be made with antibacterial activity of DMS like Co doped ZnO. In this article is found that ZnO:Co nanopowders with higher Co doping level (0.07 and 0.09wt%) exhibit good antibacterial efficiency. The magnetization curves obtained using vibrating sample magnetometer (VSM) show a sign of strong room temperature ferromagnetic behavior when the Co doping level is 0.05wt% and a weak room temperature ferromagnetic behavior Co doping level is below 0.07wt%, and also they found to exhibit antiferromagnetic and paramagnetic properties, when the Co doping levels are 0.07 and 0.09wt%, respectively, to enhance and increase the special magnetic and antibacterial property for sophisticated devices for the sustainable technologies.