Due to the increasing number of bacterial infections and the development of resistivity toward antibiotics, new materials and approaches for treatments must be urgently developed. The production of new materials should be ecologically friendly considering overall pollution with chemicals and economically acceptable and accessible to the wide population. Thus, the possibility of using biocompatible graphene quantum dots (GQDs) as an agent in photodynamic therapy was studied. First, dots were obtained using electrochemical cutting of graphite. In only one synthetic step using gamma irradiation, GQDs were doped with N atoms without any reagent. Obtained dots showed blue photoluminescence, with a diameter of 19–89 nm and optical band gap of 3.23–4.73 eV, featuring oxygen-containing, amino, and amide functional groups. Dots showed antioxidative activity; they quenched •OH at a concentration of 10 μg·mL−1, scavenged DPPH• radicals even at 5 μg·mL−1, and caused discoloration of KMnO4 at 30 μg·mL−1. Under light irradiation, dots were able to produce singlet oxygen, which remained stable for 10 min. Photoinduced effects by GQDs were studied on several bacterial strains (Listeria monocytogenes, Bacillus cereus, clinical strains of Streptococcus mutans, S. pyogenes, and S. sangunis, Pseudomonas aeruginosa, and one yeast strain Candida albicans) but antibacterial effects were not noticed.