Aureobasidin A

Background:Biofilm is known to contribute to the antifungal resistance of Candida yeasts. Aureobasidin A (AbA), a cyclic depsipeptide targeting fungal sphingolipid biosynthesis, has been shown to be effective against several Candida species.Aims:The aim of this study was to investigate Candida biofilm growth morphology, its biomass, metabolic activity, and to determine the effects of AbA on the biofilm growth.Methods:The biofilm forming ability of several clinical isolates of different Candida species from our culture collection was determined using established methods (crystal violet and XTT assays). The determination of AbA planktonic and biofilm MICs was performed based on a micro-broth dilution method. The anti-biofilm effect of AbA on Candida albicans was examined using field emission scanning electron microscope (FESEM) analysis.Results:A total of 35 (29.7%) of 118 Candida isolates were regarded as biofilm producers in this study. Candida parapsilosis was the largest producer, followed by Candida tropicalis and C. albicans. Two morphological variants of biofilms were identified in our isolates, with 48.6% of the isolates showing mainly yeast and pseudohyphae-like structures, while the remaining ones were predominantly filamentous forms. The biofilm producers were divided into two populations (low and high), based on the ability in producing biomass and their metabolic activity. Candida isolates with filamentous growth, higher biomass and metabolic activity showed lower AbA MIC50(at least fourfold), compared to those exhibiting yeast morphology, and lower biomass and metabolic activity. The observation of filament detachment and the almost complete removal of biofilm from AbA-treated C. albicans biofilm in FESEM analysis suggests an anti-biofilm effect of AbA.Conclusions:The variability in the growth characteristics of Candida biofilm cultures affects susceptibility to AbA, with higher susceptibility noted in biofilm cultures exhibiting filamentous form and high biomass/metabolic activity.

Aureobasidin A, a new antifungal antibiotic, was isolated from the culture medium of Aureobasidium pullulans R106. Aureobasidin A was a cyclic depsipeptide consisting of eight alpha-amino acid units and one hydroxy acid unit. The structures of the units were found by acid hydrolysis of the antibiotic to be 2(R)-hydroxy-3(R)-methylpentanoic acid, beta-hydroxy-N-methyl-L-valine, N-methyl-L-valine, L-proline, allo-L-isoleucine, N-methyl-L-phenylalanine, L-leucine, and L-phenyl-alanine. The sequence of the units was identified by NMR and FAB-MS of the products from the alkaline hydrolysis of aureobasidin A.

Cryptococcosis is an opportunistic fungal infection caused mainly by Cryptococcus neoformans. The aim of the present study was to evaluate the inhibitory effect of aureobasidin A on C. neoformans with special focus on its mode of action. The effect of aureobasidin A on cell membrane ergosterol content, cell wall permeability, membrane pumps activities, the total oxidant status (TOS) and melanin production was evaluated. Cytotoxicity and cell hemolysis, and laccase (LacI) and β1,2-xylosyltransferase (Cxt1p) gene expression were also evaluated. Aureobasidin A reduced melanin production and increased extracellular potassium leakage at 0.5 × MIC concentration. This peptide has no effect on fungal cell wall integrity. Cell membrane ergosterol content was decreased by 29.1% and 41.8% at 0.5 × MIC and 1 × MIC concentrations (2 and 4 µL/mL) in aureobasidin A treated samples, respectively. TOS level was significantly increased without activation of antioxidant enzymes. Lac1 gene was over-expressed (11.7-fold), while Cxt1p gene was down regulated (0.2-fold) following treatment with aureobasidin A. Overall, our results indicated that aureobasidin A inhibits C. neoformans growth by targeting different sites in fungal cells and it may be considered as a promising compound to use as an antifungal in treatment of clinical cryptococcosis.