Yeast mutants blocked in removing the methyl group of lanosterol at C-14. Separation of sterols by high-pressure liquid chromatography

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Abstract
Sterols of a nystatin resistant mutant and of the wild type parent of Saccharomyces cerevisiae were separated by a newly developed procedure involving high-pressure liquid chromatography and were identified. The mutant contained larger amounts of squalene and lanosterol than the wild type, as well as 4,14-dimethylcholesta-8,24-dien-3.beta.-ol, 4,14-dimethylergosta-8,24(28)-dien-3.beta.-ol and 14-methylergosta-8,24(28)-dien-3.beta.-ol, which were not previously found in yeast. These results indicated a block in removal of the methyl group at C-14 of lanosterol. An ergosterol requiring derivative of the mutant which carried in addition a mutation in heme biosynthesis had the same sterols as the parent, but at 1/3 the concentration. The low level of sterols may be due to requirement for a heme or cytochrome in oxygenation reactions between lanosterol and ergosterol.

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