The inactivation of spores of Penicillium oxalicum by supercritical carbon dioxide ($SC-CO_2$) was optimized by response surface methodology. The optimal inactivation conditions of 16.8 MPa, $49^{circ}C$, and 20 min were determined using ridge analysis, at which the predicted and experimental $log_{10}$ reductions were obtained as 5.74 and 6.12, respectively. The synergistic effect of a cosolvent (ethanol), which was used to modify $SC-CO_2$, on the inactivation of the fungal spores was investigated. At less severe conditions of 10 MPa and $40^{circ}C$, P. oxalicum spores of $10^7$ CFU/ml were completely inactivated within 45 min by $SC-CO_2$ modified with ethanol.