Characterization of a Saccharomyces cerevisiae Mutant, N22, Defective in Ergosterol Synthesis and Preparation of [28-14C]Ergosta-5,7-dien-3β-ol with the Mutant1

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Abstract
Analysis of sterols was made with a Saccharomyces cerevisiae mutant, N22, which was resistant to nystatin and defective in ergosterol synthesis, and with its parent strain, M10 (haploid type, methionineless, petite). The main sterol of M10 was ergosterol, whereas that of N22 was ergosta-5,7-dien-3β-ol. A small amount of ergosterol was found also in N22. This indicates that the Δ22-desaturation reaction in N22 is blocked, though not completely. Ergosta-5,8-dien-3β-ol, an unusual sterol, was detected in N22 but not in M10. Variations in the amounts and compositions of free and esterified sterols of both strains were examined during cultivation and the subsequent aerobic adaptation. The contents of free sterols, which were mostly composed of the respective main sterols of both strains, did not change markedly. In contrast, the contents and compositions of esterified sterols of both strains varied depending on the growth conditions. When the cells of both strains were aerobically adapted, the total contents of esterified sterols increased, as did, as the intermediate sterols. Upon aerobic adaptation in the presence of [methyl-14C]methionine, [28-14C]ergosta-5,7-dien-3β-ol accumulated markedly in N22 cells. Using this mutant, we devised a convenient method for preparation of the radioactive sterol in high yield.