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Literature summary for 1.1.1.201 extracted from
- Contribution of the 7beta-hydroxysteroid dehydrogenase from Ruminococcus gnavus N53 to ursodeoxycholic acid formation in the human colon (2013), J. Lipid Res., 54, 3062-3069.
Cloned(Commentary)
| 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 [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/ Products (Substrates)
| 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
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| [Ruminococcus] gnavus | A7B4V1 | isolated from human feces | - |
| [Ruminococcus] gnavus ATCC 29149 | A7B4V1 | isolated from human feces | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His6-tagged enzyme from Escherichia coli strain JM109 by nickel affinity chromatography, dialysis, and ultrafiltration | [Ruminococcus] gnavus |
Source Tissue
| 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 and Products (Substrate)
| 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
| Synonyms | Comment | Organism |
|---|---|---|
| 7beta-HSDH | - |
[Ruminococcus] gnavus |
| 7beta-hydroxysteroid dehydrogenase | - |
[Ruminococcus] gnavus |
| RUMGNA_02585 | - |
[Ruminococcus] gnavus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | [Ruminococcus] gnavus |
Temperature Stability [°C]
| 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
| 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 Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5 | 11.5 | activity range | [Ruminococcus] gnavus |
Cofactor
| 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
| 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 |
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