The Rhodococcus erythropolis SQ1 kstD3 gene was cloned, heterologously expressed and biochemically characterized as a KSTD3 (3-keto-5α-steroid Δ1-dehydrogenase). Upstream of kstD3, an ORF (open reading frame) with similarity to Δ4 KSTD (3-keto-5α-steroid Δ4-dehydrogenase) was found, tentatively designated kst4D. Biochemical analysis revealed that the Δ1 KSTD3 has a clear preference for 3-ketosteroids with a saturated A-ring, displaying highest activity on 5α-AD (5α-androstane-3,17-dione) and 5α-T (5α-testosterone; also known as 17β-hydroxy-5α-androstane-3-one). The KSTD1 and KSTD2 enzymes, on the other hand, clearly prefer (9α-hydroxy-)4-androstene-3,17-dione as substrates. Phylogenetic analysis of known and putative KSTD amino acid sequences showed that the R. erythropolis KSTD proteins cluster into four distinct groups. Interestingly, Δ1 KSTD3 from R. erythropolis SQ1 clustered with Rv3537, the only Δ1 KSTD present in Mycobacterium tuberculosis H37Rv, a protein involved in cholesterol catabolism and pathogenicity. The substrate range of heterologously expressed Rv3537 enzyme was nearly identical with that of Δ1 KSTD3, indicating that these are orthologous enzymes. The results imply that 5α-AD and 5α-T are newly identified intermediates in the cholesterol catabolic pathway, and important steroids with respect to pathogenicity.
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February 12 2008
3-Keto-5α-steroid Δ1-dehydrogenase from Rhodococcus erythropolis SQ1 and its orthologue in Mycobacterium tuberculosis H37Rv are highly specific enzymes that function in cholesterol catabolism Available to Purchase
Jan Knol;
Jan Knol
1Department of Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, PO Box 14, 9750 AA Haren, The Netherlands
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Karin Bodewits;
Karin Bodewits
1Department of Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, PO Box 14, 9750 AA Haren, The Netherlands
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Gerda I. Hessels;
Gerda I. Hessels
1Department of Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, PO Box 14, 9750 AA Haren, The Netherlands
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Lubbert Dijkhuizen;
Lubbert Dijkhuizen
1
1Department of Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, PO Box 14, 9750 AA Haren, The Netherlands
1To whom correspondence should be addressed (email [email protected]).
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Robert van der Geize
Robert van der Geize
1Department of Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, PO Box 14, 9750 AA Haren, The Netherlands
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Publisher: Portland Press Ltd
Received:
August 17 2007
Revision Received:
November 07 2007
Accepted:
November 21 2007
Accepted Manuscript online:
November 21 2007
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2008 Biochemical Society
2008
Biochem J (2008) 410 (2): 339–346.
Article history
Received:
August 17 2007
Revision Received:
November 07 2007
Accepted:
November 21 2007
Accepted Manuscript online:
November 21 2007
Citation
Jan Knol, Karin Bodewits, Gerda I. Hessels, Lubbert Dijkhuizen, Robert van der Geize; 3-Keto-5α-steroid Δ1-dehydrogenase from Rhodococcus erythropolis SQ1 and its orthologue in Mycobacterium tuberculosis H37Rv are highly specific enzymes that function in cholesterol catabolism. Biochem J 1 March 2008; 410 (2): 339–346. doi: https://doi.org/10.1042/BJ20071130
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