Cloned (Comment) | Organism |
---|---|
gene RUMGNA_02585, DNA and amino acid sequence determination and analysis, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain JM109 | [Ruminococcus] gnavus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0388 | - |
3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid | pH 6.0, 37°C, recombinant enzyme | [Ruminococcus] gnavus | |
0.974 | - |
3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid | pH 10.0, 37°C, recombinant enzyme | [Ruminococcus] gnavus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid + NADPH + H+ | [Ruminococcus] gnavus | i.e. 7-oxo-lithocholic acid | 3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+ | i.e. ursodeoxycholic acid | r | |
3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid + NADPH + H+ | [Ruminococcus] gnavus ATCC 29149 | i.e. 7-oxo-lithocholic acid | 3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+ | i.e. ursodeoxycholic acid | r | |
additional information | [Ruminococcus] gnavus | in vivo, the forward reaction that produces ursodeoxycholic acid from 7-oxo-lithocholic acid is observed to have a growth-dependent conversion rate of 90-100% after culture in anaerobic medium broth containing 1 mM 7-oxo-lithocholic acid, while the reverse reaction is undetectable. The purified 7beta-HSDH shows a kcat/Km value that is about 55fold higher for the forward reaction than for the reverse reaction, indicating that the enzyme favors the UDCA producing reaction | ? | - |
? | |
additional information | [Ruminococcus] gnavus ATCC 29149 | in vivo, the forward reaction that produces ursodeoxycholic acid from 7-oxo-lithocholic acid is observed to have a growth-dependent conversion rate of 90-100% after culture in anaerobic medium broth containing 1 mM 7-oxo-lithocholic acid, while the reverse reaction is undetectable. The purified 7beta-HSDH shows a kcat/Km value that is about 55fold higher for the forward reaction than for the reverse reaction, indicating that the enzyme favors the UDCA producing reaction | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
[Ruminococcus] gnavus | A7B4V1 | isolated from human feces | - |
[Ruminococcus] gnavus ATCC 29149 | A7B4V1 | isolated from human feces | - |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli strain JM109 by nickel affinity chromatography, dialysis, and ultrafiltration | [Ruminococcus] gnavus |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | strain N53 converts 7-oxo-lithocholic acid to ursodeoxycholic acid, and the conversion rates peaks during stationary phase, suggesting that the reductive conversion reaction is growth-dependent. In contrast, when strain N53 is cultured in the presence of ursodeoxycholic acid, 7-oxo-lithocholic acid is not detected | [Ruminococcus] gnavus | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid + NADPH + H+ | i.e. 7-oxo-lithocholic acid | [Ruminococcus] gnavus | 3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+ | i.e. ursodeoxycholic acid | r | |
3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid + NADPH + H+ | i.e 7-oxo-lithocholic acid, the forward reaction is preferred, formation of ursodeoxycholic acid | [Ruminococcus] gnavus | 3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+ | i.e. ursodeoxycholic acid | r | |
3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid + NADPH + H+ | i.e. 7-oxo-lithocholic acid | [Ruminococcus] gnavus ATCC 29149 | 3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+ | i.e. ursodeoxycholic acid | r | |
3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid + NADPH + H+ | i.e 7-oxo-lithocholic acid, the forward reaction is preferred, formation of ursodeoxycholic acid | [Ruminococcus] gnavus ATCC 29149 | 3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+ | i.e. ursodeoxycholic acid | r | |
additional information | in vivo, the forward reaction that produces ursodeoxycholic acid from 7-oxo-lithocholic acid is observed to have a growth-dependent conversion rate of 90-100% after culture in anaerobic medium broth containing 1 mM 7-oxo-lithocholic acid, while the reverse reaction is undetectable. The purified 7beta-HSDH shows a kcat/Km value that is about 55fold higher for the forward reaction than for the reverse reaction, indicating that the enzyme favors the UDCA producing reaction | [Ruminococcus] gnavus | ? | - |
? | |
additional information | in vivo, the forward reaction that produces ursodeoxycholic acid from 7-oxo-lithocholic acid is observed to have a growth-dependent conversion rate of 90-100% after culture in anaerobic medium broth containing 1 mM 7-oxo-lithocholic acid, while the reverse reaction is undetectable. The purified 7beta-HSDH shows a kcat/Km value that is about 55fold higher for the forward reaction than for the reverse reaction, indicating that the enzyme favors the UDCA producing reaction | [Ruminococcus] gnavus ATCC 29149 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
7beta-HSDH | - |
[Ruminococcus] gnavus |
7beta-hydroxysteroid dehydrogenase | - |
[Ruminococcus] gnavus |
RUMGNA_02585 | - |
[Ruminococcus] gnavus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | [Ruminococcus] gnavus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | 60 | purified recombinant His-tagged enzyme, in 100 mM MES, pH 6.0, 30 min, reductive reaction, loss of 50% activity at 50°C, and inactivation at 60°C | [Ruminococcus] gnavus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at, ursodeoxycholic acid formation | [Ruminococcus] gnavus |
10 | - |
assay at, 7-oxo-lithocholic acid formation | [Ruminococcus] gnavus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 11.5 | activity range | [Ruminococcus] gnavus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | the enzyme shows specificity toward NADPH/NADP+ against NADH/NAD+ | [Ruminococcus] gnavus | |
NADP+ | - |
[Ruminococcus] gnavus | |
NADPH | - |
[Ruminococcus] gnavus |
General Information | Comment | Organism |
---|---|---|
physiological function | the 7beta-hydroxysteroid dehydrogenase from Ruminococcus gnavus strain N53 (ATCC 29149) contributes to ursodeoxycholic acid formation in the human colon | [Ruminococcus] gnavus |