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PMID: 7766153 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Expression in Escherichia coli and characterization of a bile acid-inducible 3 alpha-hydroxysteroid dehydrogenase from Eubacterium sp. strain VPI 12708.

Current microbiology ·Vol. 30 ·No. 5 ·1995-05-00 ·Pages 259-63

Mallonee DH, Lijewski MA, Hylemon PB

Abstract

We have previously cloned and sequenced three members of a bile acid-inducible gene family from Eubacterium sp. strain VPI 12708 that encode 27,000-M(r) polypeptides. Two copies of these genes (baiA1 and baiA3) are identical, while the third copy (baiA2) encodes a polypeptide sharing 92% amino acid identity with the baiA1 and baiA3 gene products. We have overexpressed the baiA1 gene in Escherichia coli and analyzed the expressed activity. Thin-layer chromatography of 14C-labeled bile acid products from reactions using cell-free extracts revealed a 3 alpha-hydroxysteroid dehydrogenase activity for the BaiA1 protein. The BaiA1 protein could utilize both NAD+ and NADP+, and the preferred steroid substrate was the cholyl-coenzyme A conjugate rather than free cholic acid. These results show that the BaiA proteins are novel 3 alpha-hydroxysteroid dehydrogenases.

Related Genes
baiA1 baiA2 baiA3
MeSH Terms
3-Hydroxysteroid Dehydrogenases/biosynthesis,genetics,isolation & purification 3-alpha-Hydroxysteroid Dehydrogenase (B-Specific) Amino Acid Sequence Bile Acids and Salts/metabolism,pharmacology Chromatography, Thin Layer Cloning, Molecular Enzyme Induction/drug effects Escherichia coli/genetics Eubacterium/enzymology,genetics Gene Expression Genes, Bacterial Molecular Sequence Data NAD/metabolism NADP/metabolism Sequence Homology, Amino Acid Spectrophotometry
Chemicals
Bile Acids and Salts NAD NADP 3-Hydroxysteroid Dehydrogenases 3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Mallonee D H
Department of Microbiology and Immunology, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298-0678, USA.
Lijewski M A
Hylemon P B
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Article Info
Journal
Current microbiology
Abbr.
Curr Microbiol
ISSN
0343-8651
DOI
10.1007/BF00295498
Published
1995-05-00
Pages
259-63
Language
English
Region
United States
NLM ID
7808448
Grants
NIDDK NIH HHS · DK40986 · United States
NIDDK NIH HHS · F32-DK08755 · United States
NIDDK NIH HHS · P01-DK38030 · United States
External Links
PubMed source DOI: 10.1007/BF00295498 View journal details
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