Few antifungal agents are currently available for the treatment of fungal infections. Antimicrobial peptides (AMPs), which are natural molecules involved in the innate immune response of many organisms, represent a promising research method because of their broad killing activity. The aim of this study was to assess the activity of a frog AMP, [K]temporin-SHa, against some species of yeasts and moulds, and to further explore its activity against . MIC determinations were performed according to EUCAST guidelines. Next, the activity of [K]temporin-SHa against was explored using time-killing curve experiments, membrane permeabilization assays, and electron microscopy. Finally, chequerboard assays were performed to evaluate the synergy between [K]temporin-SHa and amphotericin B or fluconazole. [K]temporin-SHa was found to be active in vitro against several yeasts with MIC between 5.5 and 45 µM. [K]temporin-SHa displayed rapid fungicidal activity against (inoculum was divided into two in less than an hour and no viable colonies were recovered after 5 h) with a mechanism that could be due to membrane permeabilization. [K]temporin-SHa was synergistic with amphotericin B against (FICI = 0.303). [K]temporin-SHa could represent an additional tool to treat several species and .