In this work, the synthesis of zinc oxide (ZnO) photocatalyst from thermal decomposition of zinc acetate (ZnAc) nanoparticles obtained by supercritical antisolvent (SAS) precipitation was investigated. The optimization of calcination conditions of the SAS ZnAc was carried out, studying the effect of temperature (in the range 300–600 °C) on the production of ZnO nanoparticles. In particular, it was demonstrated that the organic residues in ZnO and its particle size, thus the specific surface area, strongly affect the photocatalytic performances. SAS micronization of ZnAc produces regular nanoparticles with a mean diameter of about 54.5 ± 11.5 nm, whereas unprocessed ZnAc is characterized by very large crystals. The experimental results evidenced that ZnAc prepared by SAS process calcined at 500 °C showed a regular nanometric structure (mean diameter: 65.0 ± 14.5 nm) and was revealed to be the best choice for the photocatalytic removal of crystal violet dye (CV). In fact, the photocatalytic activity performances of ZnO nanoparticles prepared by this route were higher with respect to that of ZnO from unprocessed ZnAc calcined at 500 °C (which is characterized by irregular tetrapods with mean size 181.1 ± 65.5 nm). The optimized photocatalyst was able to assure the complete CV decolorization in 60 min of UV irradiation time and a mineralization degree higher than 90% after 120 min of treatment time.