Characterization and regulation of the NADP-linked 7 alpha-hydroxysteroid dehydrogenase gene from Clostridium sordellii

Coleman, James P.; Hudson, L.; Adams, Margaret

doi:10.1128/jb.176.16.4865-4874.1994

Cited by 50 publications

(34 citation statements)

References 60 publications

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“…C. scindens and E. coli constitutively express 7a-HSDHs and are noninducible (69,85). Unexpectedly, the nonsubstrate DCA can induce 7a-HSDH expression in C. absonum and C. sordellii, although the reason for this induction remains unclear (70,76). Macdonald, White, and Hylemon (82) observed the expression of five soluble and two membrane polypeptides upon exposure of C. absonum to DCA and CDCA in the culture medium, although the functions of these additional polypeptides have not been determined.…”

Section: Microbial Bile Acid Hydroxysteroid Dehydrogenase(s)mentioning

confidence: 99%

“…7a/b-HSDHs are widespread among the bacteroides and clostridia as well as in E. coli and Ruminococcus species (Table 2) (54,64,(69)(70)(71)(72)(73)(74). In addition, several intestinal clostridia express both 7a-and 7b-HSDHs and have been shown to epimerize the 7a/b-hydroxy group (75,(76)(77)(78).…”

Section: Microbial Bile Acid Hydroxysteroid Dehydrogenase(s)mentioning

confidence: 99%

“…This is attributable to the intracellular location of the 7a-HSDH and the inability of the organism to take up a conjugated bile salt. The genes encoding 7a-HSDHs have been cloned from E. coli (73), C. scindens (69), and C. sordellii (70). Sequence similarity suggests that these enzymes belong to the short-chain polyol dehydrogenase family.…”

Section: Microbial Bile Acid Hydroxysteroid Dehydrogenase(s)mentioning

confidence: 99%

“…fragilis (71,79), Ba. thetaiotaomicron (74), C. scindens (80), C. sordellii (70), and E. coli (73), as well as from the soil isolates Xanthomonas maltophilia (81), C. absonum (82), and C. bifermentans (83). Interestingly, several intestinal bacteria with 7a/b-HSDH activity also possess BSH, including Ba.…”

Section: Microbial Bile Acid Hydroxysteroid Dehydrogenase(s)mentioning

confidence: 99%

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Bile salt biotransformations by human intestinal bacteria

Ridlon

Kang

Hylemon

2006

Journal of Lipid Research

2,324

2,417

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Secondary bile acids, produced solely by intestinal bacteria, can accumulate to high levels in the enterohepatic circulation of some individuals and may contribute to the pathogenesis of colon cancer, gallstones, and other gastrointestinal (GI) diseases. Bile salt hydrolysis and hydroxy group dehydrogenation reactions are carried out by a broad spectrum of intestinal anaerobic bacteria, whereas bile acid 7-dehydroxylation appears restricted to a limited number of intestinal anaerobes representing a small fraction of the total colonic flora. Microbial enzymes modifying bile salts differ between species with respect to pH optima, enzyme kinetics, substrate specificity, cellular location, and possibly physiological function. Crystallization, site-directed mutagenesis, and comparisons of protein secondary structure have provided insight into the mechanisms of several bile acid-biotransforming enzymatic reactions. Molecular cloning of genes encoding bile salt-modifying enzymes has facilitated the understanding of the genetic organization of these pathways and is a means of developing probes for the detection of bile salt-modifying bacteria. The potential exists for altering the bile acid pool by targeting key enzymes in the 7a/b-dehydroxylation pathway through the development of pharmaceuticals or sequestering bile acids biologically in probiotic bacteria, which may result in their effective removal from the host after excretion.-Ridlon, J. M., D-J. Kang, and P. B. Hylemon. Bile salt biotransformations by human intestinal bacteria. J. Lipid Res. 2006. 47: 241-259.

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Section: Microbial Bile Acid Hydroxysteroid Dehydrogenase(s)mentioning

confidence: 99%

Section: Microbial Bile Acid Hydroxysteroid Dehydrogenase(s)mentioning

confidence: 99%

Section: Microbial Bile Acid Hydroxysteroid Dehydrogenase(s)mentioning

confidence: 99%

Section: Microbial Bile Acid Hydroxysteroid Dehydrogenase(s)mentioning

confidence: 99%

See 2 more Smart Citations

Bile salt biotransformations by human intestinal bacteria

Ridlon

Kang

Hylemon

2006

Journal of Lipid Research

2,324

2,417

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“…The investigators reported that the addition of taurocholate to cecal contents from mice that had not been treated with antibiotics yielded substances that were inhibitory to C. difficile growth, while taurocholate-supplemented cecal contents from antibiotic treated mice were permissive for C. difficile growth. The authors also determined that the cecal microbiota were metabolizing taurocholate into secondary bile salts that were inhibitory for C. difficile growth, and that the presence of the enzymes responsible for converting primary bile salts into secondary bile salts were found in equivalent concentrations in both the cecum and in the feces ( Coleman et al, 1994;Berr et al, 1996;Thomas et al, 2001), an observation recently supported by meta-metabolomic profiling (Theriot et al, 2014). A recent report by Howerton et al (2013) described the use of a synthetic derivative of taurocholate, CamSA, which is a competitive inhibitor of taurocholate-induced spore germination that protected mice from challenge with a large number of C. difficile spores, further implicating the critical role of bile salts in the regulation of spore germination.…”

Section: Discussionmentioning

confidence: 99%

An in vitro culture model to study the dynamics of colonic microbiota in Syrian golden hamsters and their susceptibility to infection with Clostridium difficile

Miezeiewski¹,

Schnaufer

Muravsky

et al. 2014

The ISME Journal

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Clostridium difficile infections (CDI) are caused by colonization and growth of toxigenic strains of C. difficile in individuals whose intestinal microbiota has been perturbed, in most cases following antimicrobial therapy. Determination of the protective commensal gut community members could inform the development of treatments for CDI. Here, we utilized the lethal enterocolitis model in Syrian golden hamsters to analyze the microbiota disruption and recovery along a 20-day period following a single dose of clindamycin on day 0, inducing in vivo susceptibility to C. difficile infection. To determine susceptibility in vitro, spores of strain VPI 10463 were cultured with and without soluble hamster fecal filtrates and growth was quantified by quantitative PCR and toxin immunoassay. Fecal microbial population changes over time were tracked by 16S ribosomal RNA gene analysis via V4 sequencing and the PhyloChip assay. C. difficile culture growth and toxin production were inhibited by the presence of fecal extracts from untreated hamsters but not extracts collected 5 days post-administration of clindamycin. In vitro inhibition was re-established by day 15, which correlated with resistance of animals to lethal challenge. A substantial fecal microbiota shift in hamsters treated with antibiotics was observed, marked by significant changes across multiple phyla including Bacteroidetes and Proteobacteria. An incomplete return towards the baseline microbiome occurred by day 15 correlating with the inhibition of C. difficile growth in vitro and in vivo. These data suggest that soluble factors produced by the gut microbiota may be responsible for the suppression of C. difficile growth and toxin production.

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Efficient Synthesis of 12‐Oxochenodeoxycholic Acid Using a 12α‐Hydroxysteroid Dehydrogenase from Rhodococcus ruber

et al. 2019

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Abstract12α‐Hydroxysteroid dehydrogenase (12α‐HSDH) has the potential to convert cheap and readily available cholic acid (CA) to 12‐oxochenodeoxycholic acid (12‐oxo‐CDCA), a key precursor for chemoenzymatic synthesis of the therapeutic bile acid ursodeoxycholic acid (UDCA). In this work, a native nicotinamide adenine dinucleotide (NAD+)‐dependent 12α‐hydroxysteroid dehydrogenase (Rr12α‐HSDH) from Rhodococcus ruber was identified using a structure‐guided genome mining (SSGM) approach, which is based on the structure of cofactor pocket and the conserved nicotinamide cofactor binding motif alignment. Rr12α‐HSDH was heterologously overexpressed in Escherichia coli BL21 (DE3), purified and characterized. The purified Rr12α‐HSDH showed a high oxidative activity of 290 U mg−1protein toward CA, with a catalytic efficiency (kcat/KM) of 5.10×103 mM−1 s−1. In a preparative biotransformation (100 mL), CA (200 mM, 80 g L−1) was efficiently converted to 12‐oxo‐CDCA in 1 h, with a 85% isolated yield and a space‐time yield (STY) of up to 1632 g L−1 d−1. Furthermore, Rr12α‐HSDH was shown to be able to catalyze the oxidation of other 12α‐hydroxysteroids at high substrate loads (up to 200 mM), giving the corresponding 12‐oxo‐hydroxysteroids in 71%–85% yields, indicating the great potential of Rr12α‐HSDH as a promising biocatalyst for the synthesis of various therapeutic bile acids.magnified image

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Characterization and regulation of the NADP-linked 7 alpha-hydroxysteroid dehydrogenase gene from Clostridium sordellii

Abstract: A bile acid-inducible NADP-linked 7a-hydroxysteroid dehydrogenase (7a-HSDH) from Clostridium sordellii ATCC 9714 was purified 310-fold by ion-exchange, gel filtration, and dye-ligand affinity chromatography.

Cited by 50 publications

References 60 publications

Bile salt biotransformations by human intestinal bacteria

Bile salt biotransformations by human intestinal bacteria

An in vitro culture model to study the dynamics of colonic microbiota in Syrian golden hamsters and their susceptibility to infection with Clostridium difficile

Efficient Synthesis of 12‐Oxochenodeoxycholic Acid Using a 12α‐Hydroxysteroid Dehydrogenase from Rhodococcus ruber

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