Mesoporous TiO2 microspheres were prepared by spray pyrolysis for photocatalysis. Post modification of TiO2 by heat treatment was performed to optimize its photocatalytic performance. First, spherical TiO2 particles with mesoporous structure were synthesized at pyrolysis temperatures of 500, 600, and 700 °C. After characterization by XRD, SEM, and N2 adsorption, a sample prepared at 500 °C was found to possess desirable properties for photocatalytic performance through post-modification. In methylene blue degradation, mesoporous TiO2 microspheres synthesized at 500 °C outperformed other microspheres. Furthermore, samples obtained by spray pyrolysis at 500 °C were calcined at various temperatures as a post-modification process. The sample calcined at 350 °C showed improved photocatalytic activity due to optimal anatase crystallinity and surface area.