The interaction of the first-generation quinolone antibacterial drug oxolinic acid (Hoxo) with the dioxomolybdenum(VI) and dioxouranim(VI) ions leads to the formation of the neutral mononuclear complexes MoO2(oxo)2 and UO2(oxo)2, respectively. The structure of the complexes has been characterized physicochemically and spectroscopically. The lowest energy model structure of the complexes has been determined with molecular modeling calculations. The antimicrobial activity of the complexes has been evaluated against three different microorganisms. The interaction of the complexes with calf-thymus DNA has been investigated with electronic and circular dichroism spectroscopies.