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Aspergillus fumigatus intrinsic fluconazole resistance has been demonstrated to be linked to theCYP51A gene, although the precise molecular mechanism has not been elucidated yet. Comparisons betweenA. fumigatus Cyp51Ap andCandida albicans Erg11p sequences showed differences in amino acid residues already associated with fluconazole resistance inC. albicans . The aim of this study was to analyze the role of the natural polymorphism I301 inAspergillus fumigatus Cyp51Ap in the intrinsic fluconazole resistance phenotype of this pathogen. The I301 residue inA. fumigatus Cyp51Ap was replaced with a threonine (analogue to T315 atCandida albicans fluconazole-susceptible Erg11p) by changing one single nucleotide in theCYP51A gene. Also, aCYP51A knockout strain was obtained using the same parental strain. Both mutants' antifungal susceptibilities were tested. The I301T mutant exhibited a lower level of resistance to fluconazole (MIC, 20 μg/ml) than the parental strain (MIC, 640 μg/ml), while no changes in MIC were observed for other azole- and non-azole-based drugs. These data strongly implicate theA. fumigatus Cyp51Ap I301 residue in the intrinsic resistance to fluconazole.

Aspergillus fumigatus Intrinsic Fluconazole Resistance Is Due to the Naturally Occurring T301I Substitution in Cyp51Ap