Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Clostridium sardiniense |
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 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 | 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 (Comment) | Organism |
---|---|
Ni-IDA affinity column chromatography | Clostridium sardiniense |
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 | 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 | Comment | Organism |
---|---|---|
? | x * 31000, SDS-PAGE | Clostridium sardiniense |
Synonyms | Comment | Organism |
---|---|---|
NADP+-dependent 7alpha-HSDH | - |
Clostridium sardiniense |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
- |
Clostridium sardiniense |
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 | Comment | Organism | Structure |
---|---|---|---|
NADP+ | dependent on | Clostridium sardiniense |
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 |