Pure and manganese-doped titanium dioxide nanoparticles (MnTiO2-NPs) were synthesized by the defect-oriented hydrothermal approach. The synthesized material was then characterized by X-ray diffraction (XRD), Scanning electron microscopy (SEM), Energy dispersive X-ray spectroscopy (EDX), and UV–visible spectroscopy (UV–Vis). The agar well diffusion method assessed the antibacterial efficiency of TiO2 and MnTiO2-NPs against E. coli and S. aureus. Zone of inhibition (ZOI) formed by pure TiO2 was observed as 12 mm and 11.5 mm against E. coli and S. aureus, while for MnTiO2-NPs it was observed as 19 mm (E. coli) and 21 mm (S. aureus). The concentration of synthesized nanoparticles (10 mg/ml, and 20 mg/ml) was used for antibacterial studies. The efficacy of the pure and MnTiO2-NPs as an active photocatalyst for the degradation of methylene blue (MB) dye was also assessed using a UV light. It was observed that the photodegradation efficiency of 1 g of MnTiO2-NPs was higher than the same amount of pure TiO2. The results suggest that the photocatalyst concentration directly impacts the photodegradation of MB dye. The pH value was found to influence the photodegradation of MB dye at higher pH values. Based on the obtained results, MnTiO2-NPs were observed as a promising agent for microbial resistance and water remediation.