Antifungal and Antibiofilm Activities and the Mechanism of Action of Repeating Lysine-Tryptophan Peptides against .

The rapid increase in the emergence of antifungal-resistant strains is becoming a serious health concern. Because antimicrobial peptides (AMPs) may provide a potential alternative to conventional antifungal agents, we have synthesized a series of peptides with a varying number of lysine and tryptophan repeats (KW-NH). The antifungal activity of these peptides increased with peptide length, but only the longest KW peptide displayed cytotoxicity towards a human keratinocyte cell line. The KW and KW peptides exhibited strong antifungal activity against , even under conditions of high-salt and acidic pH, or the addition of fungal cell wall components. Moreover, KW inhibited biofilm formation by a fluconazole-resistant strain. Circular dichroism and fluorescence spectroscopy indicated that fungal liposomes could interact with the longer peptides but that they did not release the fluorescent dye calcein. Subsequently, fluorescence assays with different dyes revealed that KW did not disrupt the membrane integrity of intact fungal cells. Scanning electron microscopy showed no changes in fungal morphology, while laser-scanning confocal microscopy indicated that KW can localize into the cytosol of . Gel retardation assays revealed that KW can bind to fungal RNA as a potential intracellular target. Taken together, our data indicate that KW can inhibit cellular functions by binding to RNA and DNA after it has been translocated into the cell, resulting in the eradication of .