Cytochrome p-450 reductase screen for ergosterol biosynthesis inhibitors

Abstract

A method for screening for potential fungicides identifies biochemical or chemical agents that inhibit the formation of the fungal sterol ergosterol by inhibiting cytochrome P450 reductase or reductase related enzymes or proteins involved in its biosynthesis. Test samples are incubated in two yeast strain cultures, and the extent of growth inhibition is compared between them. One culture is supersensitive to cytochrome P450 reductase inhibitors; the yeast strain typically contains a disrupted or deleted cytochrome P450 reductase gene. The other culture is less sensitive to the compounds, typically exhibiting enhanced permeability such as that observed in erg6 mutants. In preferred embodiments, a known cytochrome P450 reductase related electron-transfer protein inhibitor such as keto-conazole, miconazole, dinaconazole or econazole is employed as a positive control. Preferred embodiments employ solidified media, and test samples are applied to disks or wells. Actives are identified by the production of a zone differential of approximately 8 mm or greater on the yeast culture lawns.