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  • Title: Preparation of 14C-labeled and unlabeled sterol intermediates from yeast using metabolic inhibitors.
    Author: Nakanishi S, Nishino T, Yabusaki Y, Fujisaki S, Katsuki H.
    Journal: J Biochem; 1984 Dec; 96(6):1665-72. PubMed ID: 6397465.
    Abstract:
    The method for the preparation of zymosterol was improved (13 mg of zymosterol/g dry cells) by the aerobic adaptation of the cells in the presence of 1 mM DL-ethionine. Lanosterol was also found to accumulate (5.0 mg/g dry cells) when the cells were adapted aerobically in the presence of 10(-4) M buthiobate. Pure lanosterol could be obtained by separation of the unsaponifiable lipids on TLC. Pure [14C]lanosterol with a high specific radioactivity (56 Ci/mol) could be prepared by incubation of the desiccated cells with [14C]isopentenyl pyrophosphate, cofactors such as ATP and NADPH-generating system, and buthiobate in phosphate buffer. The method using desiccated cells may also be applicable to the preparation of other radioactive sterol intermediates.
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