TiO 2 nanoparticles were synthesized at room temperature by chemical precipitation method and were then heated at 120, 300, 600 and 900 °C temperatures. The phase transition and crystallite size variation were determined by X-rays diffraction (XRD) analysis. The surface area, pore volume and pore size were measured using Brunauer-Emmet-Teller (BET) and Barrett-Joyner-Halenda (BJH) methods. The optical activity of heat treated and non-heat treated samples were carried out by diffuse reflectance (DR) spectroscopy. Four different methods were used to calculate band gap energy. The results obtained from thermogravimetric and differential thermal gravimetric (TG/TDG) analyses and Fourier transform infra-red (FTIR) spectroscopy agreed with each other. Agar well diffusion method has been applied to explore the antibacterial activity of nanoparticles against different bacterial strains such as Bacillus subtilis, Staphylococcus Aureus, Escherichia coli and Pseudomonas Aeruginosa. It was observed that TiO 2 nanoparticles heated at 120 °C displayed maximum antibacterial activity while those heated at higher temperature showed no activity against the examined bacteria.