Sterol 24(28) methylene reductase in Saccharomyces cerevisiae Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/cn69m7844

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  • A cell free extract from Saccharomyces cerevisiae containing the enzyme that causes reduction of the 24(28)methylene group on the side chain of ergosterol has been studied. Optimal conditions for the 24(28)methylene reductase were obtained. The enzyme activity was increased in cells grown with ethanol as the substrate. The Km was calculated as 10 μM. Within the limits of the assay, enzymatic activity was not present in ergostatetraene accumulating mutants that were tested. The enzyme was competitively inhibited when tested against a naturally occurring azasterol. The dissociation constant (KI) was calculated as 17 μM. The enzyme reaction is not reversible under the conditions provided by the assay.
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