Defect-engineering of TiO2 can have a major impact on its photocatalytic properties for the degradation of persisting and non-biodegradable pollutants. Herein, a series of intrinsic and extrinsic defects are induced by post annealing of crystalline TiO2 under different reducing atmospheres. A detailed optoelectronic characterization sheds light on the key characteristics of the defect-engineered TiO2 nanopowders that are linked to the photocatalytic performance of the prepared photocatalysts. The photodegradation of a model dye, malachite green, as well as the inactivation of bacterial endospores of the Geobacillus stearothermophilus species were studied in the presence of the developed catalysts under visible light illumination. Our results indicate that a combination of certain defects is necessary for the improvement of the photocatalytic process for water purification and disinfection under visible light.