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• W2040532413 abstract "A gram-positive, rod-shaped anaerobe (strain F-6) was isolated from soil. This organism was identified by cellular morphology as well as fermentative and biochemical data as Clostridium bifermentans. Strain F-6 formed 7-ketolithocholic acid from chenodeoxycholic acid and 7-ketodeoxycholic acid from cholic acid in whole cell cultures, but did not transform deoxycholic acid, ursodeoxycholic acid, or ursocholic acid. This reaction is reversible. The structures of 7-ketolithocholic acid and 7-ketodeoxycholic acid were verified by mass spectroscopy and by thin-layer chromatography using Komarowsky's spray reagent. When incubated with the strain F-6 glycine and taurine conjugates of the primary bile acids were partially hydrolyzed and transformed to 7-keto products. Optimal yields of 7-ketolithocholic acid and 7-ketodeoxycholic acid were obtained after 78 h of incubation. Culture pH changed with time and was characterized by an initial drop (1.1 pH units) and a gradual increase back to the starting pH (7.3). Corroborating these observations, an inducible, NADP-dependent, 7α-hydroxysteroid dehydrogenase was demonstrated in cell extracts of strain F-6. A trace of NAD-dependent 7α-hydroxysteroid dehydrogenase was also found. A substantial increase in the specific activity of the NADP-dependent 7α-hydroxysteroid dehydrogenase was observed when either 7-ketolithocholic acid, chenodeoxycholic acid, or deoxycholic acid was included in the growth medium. Optimal induction of the NADP-dependent 7α-hydroxysteroid dehydrogenase was achieved with 0.3–0.4 mM 7-ketolithocholic acid. Production of the enzyme(s) was optimal at 6–8 h of growth and the 7α-hydroxysteroid dehydrogenases had a pH optimum of approximately 11. The 7α-hydroxysteroid dehydrogenase from strain F-6 was purified 12-fold by triazine dye affinity chromatography with reactive blue 2 (Cibacron blue) agarose (95% yield). It was successfully lyophilized into a stable powder form." @default.
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• W2040532413 date "1987-08-01" @default.
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• W2040532413 title "Oxidation of primary bile acids by a 7α-hydroxysteroid dehydrogenase elaborating <i>Clostridium bifermentans</i> soil isolate" @default.
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• W2040532413 doi "https://doi.org/10.1139/m87-116" @default.
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