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Literature summary extracted from
- Asp-99 donates a hydrogen bond not to Tyr-14 but to the steroid directly in the catalytic mechanism of DELTA 5-3-ketosteroid isomerase from Pseudomonas putida biotype B (2000), Biochemistry, 39, 903-909.
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 5.3.3.1 | mutant enzyme D99E/D38N complexed with equilenin, an intermediate analogue, crystals of the complexes are grown from 1.1 M ammonium acetate and 0.1 M sodium acetate, pH 4.6, by the hanging drop vapor diffusion method at 22°C. The resulting crystals have C2 space group symmetry with unit cell dimensions of a = 89.04 A, b = 72.42 A, c = 51.24 A and beta = 90.9° | Pseudomonas putida |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 5.3.3.1 | D99E | turnover-number for 5-androstene-3,17-dione is 68.1% of the turnover number of the wild-type enzyme, the Km-value is 1.22fold higher than the Km-value of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | D99L | turnover-number for 5-androstene-3,17-dione is 125fold lower than the turnover number of the wild-type enzyme, the Km-value is 1.95fold lower than the Km-value of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | Y14F | turnover-number for 5-androstene-3,17-dione is less than 1% of the turnover number of the wild-type enzyme, the Km-value is 1.87fold lower than the Km-value of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | Y14F/D99E | turnover-number for 5-androstene-3,17-dione is less than 1% of the turnover number of the wild-type enzyme, the Km-value is 1.05fold lower than the Km-value of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | Y14F/D99L | turnover-number for 5-androstene-3,17-dione is less than 1% of the turnover number of the wild-type enzyme, the Km-value is 1.95fold higher than the Km-value of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | Y14F/D99N | turnover-number for 5-androstene-3,17-dione is less than 1% of the turnover number of the wild-type enzyme, the Km-value is 1.06fold lower than the Km-value of the wild-type enzyme | Pseudomonas putida |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.3.3.1 | 19-nortestosterone | - |
Pseudomonas putida |  |
| 5.3.3.1 | equilenin | - |
Pseudomonas putida | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.3.3.1 | 0.0269 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme Y14F | Pseudomonas putida | |
| 5.3.3.1 | 0.0475 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme Y14F/D99N | Pseudomonas putida | |
| 5.3.3.1 | 0.0481 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme Y14F/D99E | Pseudomonas putida | |
| 5.3.3.1 | 0.0503 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, wild-type enzyme | Pseudomonas putida | |
| 5.3.3.1 | 0.0616 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme D99E | Pseudomonas putida | |
| 5.3.3.1 | 0.0982 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme Y14F/D99L | Pseudomonas putida | |
| 5.3.3.1 | 0.258 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme D99L | Pseudomonas putida | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 5.3.3.1 | Pseudomonas putida | - |
- |
- |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.3.3.1 | 5-Androstene-3,17-dione | - |
Pseudomonas putida | 4-Androstene-3,17-dione | - |
? | |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.3.3.1 | 0.67 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme Y14F/D99L | Pseudomonas putida | |
| 5.3.3.1 | 0.7 | 1 | 5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme Y14F/D99N | Pseudomonas putida | |
| 5.3.3.1 | 0.71 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme Y14F/D99N | Pseudomonas putida | |
| 5.3.3.1 | 3 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme Y14F/D99E | Pseudomonas putida | |
| 5.3.3.1 | 6.08 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme Y14F/D99L | Pseudomonas putida | |
| 5.3.3.1 | 11.5 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme Y14F | Pseudomonas putida | |
| 5.3.3.1 | 220 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme D99L | Pseudomonas putida | |
| 5.3.3.1 | 19000 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme D99E | Pseudomonas putida | |
| 5.3.3.1 | 27900 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, wild-type enzyme | Pseudomonas putida | |
Ki Value [mM]
| EC Number | Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.3.3.1 | additional information | - |
additional information | Ki-values for equilenin and 19-nortestosterone, wild-type enzyme and mutant enzyme D99E | Pseudomonas putida |
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