Effect of pH on bile salt degradation by mixed fecal cultures

Stoll specimens from 3 healthy volunteers were cultured under an-aerobic conditions in brain heart infusion broth with and without the addition of cholate, deoxycholate of chenodeoxycholate. The initial pH of the medium was adjusted to 5.5, 6.3, 7.3 (unadjusted), 8.0, and 9.0. Cell-free extracts prepared from the resulting bacterial growth contained increased levels of NAD- and NADP-dependent 3alpha-, 7alpha-, and 12alpha-hydroxysteroid oxidoreductases when the initial pH was 8.0 or 9.0 and depressed levels of these activities when the initial pH was 5.5 or 6.3 (as compared to control values obtained at 7.3). At pH 5,5 all activities except NAD-dependent 7alpha-hydroxysteroid oxidoreductase were absent. A powerful selective effect was imposed on NAD-dependent 7alpha-hydroxysteroid oxidoreductase when deoxycholate or chenodeoxycholate were incorporated into or chenodeoxycholate were incorporated into the medium. Thin-layer chromatography of either extracts of cholate-containing, acidified spent bacterial medium showed alkaline or neutral (optimal at pH 8). The precent hydroxyl group estimations at the 3alpha-, 7alpha-, and 12alpha-positions revealed an increase in disappearance of OH groups at all three positions with increasing initial pH value. The order of extent of bioconversion was 7alpha-OH greater than 3alpha-OH; at pH 8 AND 9, approximately 90% 7alpha-OH bioconversion was observed. Spent bacterial media and a number of commercial secondary bile salts were all negative in the Ames' assay for mutagenicity.