Cloned (Comment) | Organism |
---|---|
overoverexpression with an N-terminal His-tag in Escherichia coli | Pseudomonas sp. B-0831 |
Protein Variants | Comment | Organism |
---|---|---|
K157A | the NADH binding affinity of K157A mutant is much lower than that of the wild-type, mainly due to loss of a hydrogen bond | Pseudomonas sp. B-0831 |
S114A | mutant shows higher Km and lower kcat values in both oxidation and reduction reactions | Pseudomonas sp. B-0831 |
S114A/Y153F | double mutation results in a significant decrease in kcat relative to the single mutant Y153F | Pseudomonas sp. B-0831 |
Y153F | mutant shows higher Km and lower kcat values in both oxidation and reduction reactions. Loss of hydrogen bonding with NADH upon the Y153F mutation results in increased enthalpy change | Pseudomonas sp. B-0831 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
51 | - |
dehydrocholate | 37°C, pH 8.0, wild-type enzyme | Pseudomonas sp. B-0831 | |
56 | - |
cholate | 37°C, pH 8.0, wild-type enzyme | Pseudomonas sp. B-0831 | |
78 | - |
cholate | 37°C, pH 8.0, mutant enzyme S114A/Y153F | Pseudomonas sp. B-0831 | |
96 | - |
cholate | 37°C, pH 8.0, mutant enzyme K157A | Pseudomonas sp. B-0831 | |
239 | - |
dehydrocholate | 37°C, pH 8.0, mutant enzyme S114A | Pseudomonas sp. B-0831 | |
468 | - |
cholate | 37°C, pH 8.0, mutant enzyme Y153F | Pseudomonas sp. B-0831 | |
586 | - |
cholate | 37°C, pH 8.0, mutant enzyme S114A | Pseudomonas sp. B-0831 | |
2489 | - |
dehydrocholate | 37°C, pH 8.0, mutant enzyme Y153F | Pseudomonas sp. B-0831 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. B-0831 | Q59718 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Pseudomonas sp. B-0831 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cholate + NAD+ | - |
Pseudomonas sp. B-0831 | 7alpha,12alpha-dihydroxy-5-beta-cholan-3-one-24-oic acid + NADH + H+ | - |
? | |
dehydrocholate + NADH + H+ | - |
Pseudomonas sp. B-0831 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
3alpha-hydroxysteroid dehydrogenase | - |
Pseudomonas sp. B-0831 |
Ps3alphaHSD | - |
Pseudomonas sp. B-0831 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.045 | - |
cholate | 37°C, pH 8.0, mutant enzyme K157A | Pseudomonas sp. B-0831 | |
0.12 | - |
cholate | 37°C, pH 8.0, mutant enzyme S114A/Y153F | Pseudomonas sp. B-0831 | |
0.62 | - |
cholate | 37°C, pH 8.0, mutant enzyme Y153F | Pseudomonas sp. B-0831 | |
0.8 | - |
cholate | 37°C, pH 8.0, mutant enzyme S114A | Pseudomonas sp. B-0831 | |
0.89 | - |
dehydrocholate | 37°C, pH 8.0, mutant enzyme S114A | Pseudomonas sp. B-0831 | |
1.1 | - |
dehydrocholate | 37°C, pH 8.0, mutant enzyme Y153F | Pseudomonas sp. B-0831 | |
23 | - |
cholate | 37°C, pH 8.0, wild-type enzyme | Pseudomonas sp. B-0831 | |
48 | - |
dehydrocholate | 37°C, pH 8.0, wild-type enzyme | Pseudomonas sp. B-0831 |