By using cell-free preparations of rat liver it was shown that the removal of the 14alpha-methyl group (C-32) of steroids containing either a delta7(8) or a delta8(9) double bond is attended exclusively by the formation of the corresponding 7,14- and 8,14-dienes respectively (structures of types III and VIII). Cumulative evidence from a variety of experimental approaches had led to the deduction that delta8(14)-steroids are not involved as intermediates on the major pathway of cholesterol biosynthesis. The metabolism of [32-3H]lanost-7-ene-3beta,32-diol (structure of type I) results in the formation of radioactive formic acid, no labelled formaldehyde being formed. By using appropriately labelled species of the compound (I) it was found that the release of formic acid and the formation of 4,4-dimethylcholesta-7,14-dien-3beta-ol (strurcture of type III) were closely linked processes, and that in the conversion of compound (I) into compound (III), 3-beta-hydroxylanost-7-en-32-al (II) is an obligatory intermediate. Both the conversion of lanost-7-ene-3beta,32-diol (I) into 3beta-hydroxylanost-7-en-32-al (II) and the further metabolism of the latter (II) to 4,4-dimethylcholesta-7,14-dien-3beta-ol (III) exhibited a requirement for NADPH and O2. This suggests that the oxidation of the 32-hydroxy group of compound (I) to the aldehyde group of compound (II) does not occur by the conventional alcohol dehydrogenase type of reaction, but may proceed by a novel mechanism involving the intermediacy of a gem-diol. A detailed overall pathway for the 14alpha-demethylation in cholesterol biosynthesis is considered, and proposals about the mechanism of individual steps in the pathway are made.

It is shown that the removal of the 32-carbon atom in cholesterol biosynthesis occurs a t the oxidation state of an aldehyde resulting in the release of formic acid.

No abstract

If the biological conversion of cholest-7-en-3beta-ol (I) into cholesterol (IV) occurred thorugh the intermediacy of cholest-7-ene-3beta,5alpha-diol (II) then the factor(s) adversely affecting the convwesion of the 5alpha-hydroxy sterol (II) into cholesterol must at least equally adversely affect the formation of cholesterol from cholest-7-en-3beta-ol. By using partial denaturation techniquws and dual-labelled precursors it was shown that the enzyme system responsible for the conversion of the 5alpha-hydroxy sterol (II) into cholesterol denatured faster than that for the corresponding conversion from cholest-7-en-3beta-ol (I).