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Literature summary for 1.1.1.159 extracted from
- Enhanced activity and substrate tolerance of 7alpha-hydroxysteroid dehydrogenase by directed evolution for 7-ketolithocholic acid production (2019), Appl. Microbiol. Biotechnol., 103, 2665-2674 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli BL21(DE3) cells | Clostridium sardiniense |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| C260S | the mutant shows 2fold higher specific activity as compared to the wild type enzyme | Clostridium sardiniense |
| L26M | the mutant shows 1.26fold higher specific activity as compared to the wild type enzyme | Clostridium sardiniense |
| Q255L | the mutant shows 4.2fold higher specific activity as compared to the wild type enzyme | Clostridium sardiniense |
| Q255L/C260S | the mutant shows 6.5fold higher specific activity and exhibits 10fold higher and 14fold higher catalytic efficiencies toward chenodeoxycholic acid and NADP+, respectively, as compared to the wild type enzyme. The mutant also displays significantly enhanced tolerance in the presence of high concentrations of substrate compared to the wild type | Clostridium sardiniense |
| Q255R | the mutant shows 1.3fold higher specific activity as compared to the wild type enzyme | Clostridium sardiniense |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0601 | - |
chenodeoxycholic acid | mutant enzyme Q255L/C260S, at pH 8.0 and 25°C | Clostridium sardiniense |  |
| 0.0661 | - |
chenodeoxycholic acid | wild type enzyme, at pH 8.0 and 25°C | Clostridium sardiniense | |
| 0.1094 | - |
NADP+ | mutant enzyme Q255L/C260S, at pH 8.0 and 25°C | Clostridium sardiniense | |
| 0.3116 | - |
NADP+ | wild type enzyme, at pH 8.0 and 25°C | Clostridium sardiniense | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Clostridium sardiniense | G9FRD7 | formerly known as Clostridium sardiniense strain DSM599 | - |
| Clostridium sardiniense DSM 1599 | G9FRD7 | formerly known as Clostridium sardiniense strain DSM599 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| Ni-IDA affinity column chromatography | Clostridium sardiniense |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 5.7 | - |
wild type enzyme, at pH 8.0 and 25°C | Clostridium sardiniense |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| chenodeoxycholic acid + NADP+ | - |
Clostridium sardiniense | 7-ketolithocholic acid + NADPH + H+ | - |
r | |
| chenodeoxycholic acid + NADP+ | - |
Clostridium sardiniense DSM 1599 | 7-ketolithocholic acid + NADPH + H+ | - |
r | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 31000, SDS-PAGE | Clostridium sardiniense |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| NADP+-dependent 7alpha-HSDH | - |
Clostridium sardiniense |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
- |
Clostridium sardiniense |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 2.34 | - |
chenodeoxycholic acid | wild type enzyme, at pH 8.0 and 25°C | Clostridium sardiniense | |
| 3.66 | - |
NADP+ | wild type enzyme, at pH 8.0 and 25°C | Clostridium sardiniense | |
| 18.45 | - |
NADP+ | mutant enzyme Q255L/C260S, at pH 8.0 and 25°C | Clostridium sardiniense | |
| 21 | - |
chenodeoxycholic acid | mutant enzyme Q255L/C260S, at pH 8.0 and 25°C | Clostridium sardiniense | |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADP+ | dependent on | Clostridium sardiniense | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 11.8 | - |
NADP+ | wild type enzyme, at pH 8.0 and 25°C | Clostridium sardiniense | |
| 35.4 | - |
chenodeoxycholic acid | wild type enzyme, at pH 8.0 and 25°C | Clostridium sardiniense | |
| 168.6 | - |
NADP+ | mutant enzyme Q255L/C260S, at pH 8.0 and 25°C | Clostridium sardiniense | |
| 349.4 | - |
chenodeoxycholic acid | mutant enzyme Q255L/C260S, at pH 8.0 and 25°C | Clostridium sardiniense | |
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Release 2023.1 (January 2023)
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