| Oxidation of primary bile acids by a 7 alpha-hydroxysteroid dehydrogenase elaborating Clostridium bifermentans soil isolate. | |
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MedLine Citation:
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PMID: 3480039 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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, 7 alpha-hydroxysteroid dehydrogenase was demonstrated in cell extracts of strain F-6. A trace of NAD-dependent 7 alpha-hydroxysteroid dehydrogenase was also found. A substantial increase in the specific activity of the NADP-dependent 7 alpha-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 7 alpha-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 7 alpha-hydroxysteroid dehydrogenases had a pH optimum of approximately 11.(ABSTRACT TRUNCATED AT 250 WORDS) |
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Authors:
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J D Sutherland; C N Williams; D M Hutchison; L V Holdeman |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Canadian journal of microbiology Volume: 33 ISSN: 0008-4166 ISO Abbreviation: Can. J. Microbiol. Publication Date: 1987 Aug |
Date Detail:
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Created Date: 1988-01-25 Completed Date: 1988-01-25 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0372707 Medline TA: Can J Microbiol Country: CANADA |
Other Details:
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Languages: eng Pagination: 663-9 Citation Subset: IM |
Affiliation:
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Department of Medicine, Dalhousie University, Halifax, NS, Canada. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Bile Acids and Salts
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metabolism* Biotransformation Chromatography, Affinity Clostridium / enzymology*, growth & development Hydrogen-Ion Concentration Hydroxysteroid Dehydrogenases / isolation & purification, metabolism* Kinetics Oxidation-Reduction Soil Microbiology* Temperature |
| Chemical | |
Reg. No./Substance:
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0/Bile Acids and Salts; EC 1.1.-/Hydroxysteroid Dehydrogenases; EC 1.1.1.159/7 alpha-hydroxysteroid dehydrogenase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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