The conversion of cholest-7-en-3beta-ol into cholesterol. General comments on the mechanism of the introduction of double bonds in enzymic reactions

Abstract

Convenient syntheses of 6beta-tritiated Delta(7)-cholestenol and 3alpha-tritiated Delta(7)-cholestene-3beta,5alpha-diol are described. It was shown that the conversion of 6beta-tritiated Delta(7)-cholestenol into cholesterol is accompanied by the complete retention of label. It was unambiguously established that the overall reaction leading to the introduction of the double bond in the 5,6-position in cholesterol occurs via a cis-elimination involving the 5alpha- and 6alpha-hydrogen atoms and that during this process the 6beta-hydrogen atom remains completely undisturbed. Metabolic studies with 3alpha-tritiated Delta(7)-cholestene-3beta,5alpha-diol revealed that under anaerobic conditions the compound is not converted into cholesterol. This observation, coupled with the previous work of Slaytor & Bloch (1965), is interpreted to exclude a hydroxylation-dehydration mechanism for the origin of the 5,6-double bond in cholesterol. It was also shown that under aerobic conditions 3alpha-tritiated Delta(7)-cholestene-3beta,5alpha-diol is efficiently converted into cholesterol and that this conversion occurs through the intermediacy of 7-dehydrocholesterol. Cumulative experimental evidence presented in this paper and elsewhere is used to suggest that the 5,6-double bond in cholesterol originates through an oxygen-dependent dehydrogenation process and a hypothetical mechanism for this and related reactions is outlined.