Background: The microwave energy is an efficient disinfection method; however, it can generate high temperatures that can result in distortion of the dentures. objectives: To evaluate whether the addition of an enzymatic cleanser to microwave disinfection regimen would disinfect dentures with shorter irradiation time. Materials and methods: Seven resin discs colonized with Candida albicans biofilm were placed on the palatal surface of sterile dentures to be randomly assigned to the following treatments: immersion in distilled water for 3 min with 0 (DW), 1 (DW + M1), 2 (DW + M2), or 3 min (DW + M3) of microwave irradiation; or immersion in denture cleanser for 3 min with 0 (DC), 1 (DC + M1), 2 (DC + M2) or 3 min (DC + M3) of irradiation. After the treatments, the viable cells were counted by a blinded examiner. The temperature was measured immediately after irradiation. The data were analyzed by ANOVA and Tukey post hoc tests ( = 0.05). Results: No viable cells were found after DC + M2, DC + M3, and DW + M3 treatments, of which DC + M2 achieved the lowest temperature. No significant difference was found between the effectiveness of DW, DW + M1 and DC treatments (p > 0.05). Conclusion: Within the limits of this study, the association of a denture cleanser and microwave energy is efficient to disinfect dentures in lower irradiation time and temperature.3012631