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Literature summary extracted from
- Roles of active site aromatic residues in catalysis by ketosteroid isomerase from Pseudomonas putida biotype B (1999), Biochemistry, 38, 13810-13819.
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 5.3.3.1 | crystal structure of mutant enzyme F82A is determined to 2.1 A resolution. Crystals are grown in a solution containing 1.0 M sodium acetate and 0.1 M ammonioum acetate, pH 4.6, by the hanging drop method of vapor diffusion at 22 C. The crystals belong to the space group c2221 with unit cell dimensions of a = 36.24 A, b = 96.13 A and c = 74.30 A | Pseudomonas putida |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 5.3.3.1 | F54A | turnover number for 5-androstene-3,17-dione is 17.1% of the turnover number for the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is 15.3% of the turnover number of the wild-type enzyme,the KM-value for 5-androstene-3,17-dione is 116% of the KM-value of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 233% of the KM-value of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | F54L | turnover number for 5-androstene-3,17-dione is 41.9% of the turnover number of the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is 38.7% of the turnover number of the wild-type enzyme, the KM-value for 5-androstene-3,17-dione is 154% of the KM-value of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 89.3% of the KM-value of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | F82A | turnover number for 5-androstene-3,17-dione is 2.5% of the turnover number of the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is 2.3% of the turnover number of the wild-type enzyme,the KM-value for 5-androstene-3,17-dione is 29.9% of the KM-value of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 29.8% of the KM-value of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | F82L | turnover number for 5-androstene-3,17-dione is 12% of the turnover number of the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is 67.6% of the turnover number of the wild-type enzyme, the KM-value for 5-androstene-3,17-dione is 108% of the KM-value of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 130% of the KM-value of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | W116A | turnover number for 5-androstene-3,17-dione is 1.5% of the turnover number of the wild-type enzyme, turnover number for 5(10)-estrene-3,17-dione is below 12%% of the turnover number of the wild-type enzyme, the KM-value for 5-androstene-3,17-dione is 348% of the KM-value of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | W116F | turnover number for 5-androstene-3,17-dione is 22.4% of the turnover number of the wild-type enzyme, the KM-value for 5(10)-estrene-3,17-dione is 299% of the KM-value of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | W116Y | turnover number for 5-androstene-3,17-dione is 21% of the turnover number of the wild-type enzyme | Pseudomonas putida |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.3.3.1 | 0.0149 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82A | Pseudomonas putida |  |
| 5.3.3.1 | 0.0328 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82A | Pseudomonas putida | |
| 5.3.3.1 | 0.0499 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, wild-type enzyme | Pseudomonas putida | |
| 5.3.3.1 | 0.0539 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82L | Pseudomonas putida | |
| 5.3.3.1 | 0.0579 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54A | Pseudomonas putida | |
| 5.3.3.1 | 0.0651 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82L | Pseudomonas putida | |
| 5.3.3.1 | 0.077 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54L | Pseudomonas putida | |
| 5.3.3.1 | 0.0982 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54L | Pseudomonas putida | |
| 5.3.3.1 | 0.11 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, wild-type enzyme | Pseudomonas putida | |
| 5.3.3.1 | 0.149 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme W116F | Pseudomonas putida | |
| 5.3.3.1 | 0.174 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme W116A | Pseudomonas putida | |
| 5.3.3.1 | 0.256 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54A | Pseudomonas putida | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 5.3.3.1 | Pseudomonas putida | - |
biotype B | - |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.3.3.1 | 5(10)-estrene-3,17-dione | - |
Pseudomonas putida | estr-4-en-3,17-dione | - |
? | |
| 5.3.3.1 | 5-Androstene-3,17-dione | - |
Pseudomonas putida | 4-Androstene-3,17-dione | - |
? | |
| 5.3.3.1 | additional information | the aromatic residues Phe54, Phe82 and Trp116 in the hydrophobic substrate-binding pocket of the enzyme contribute differentially to steroid species including substrate, intermediate and product | Pseudomonas putida | ? | - |
? | |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.3.3.1 | 0.25 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82A | Pseudomonas putida | |
| 5.3.3.1 | 1.7 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54A | Pseudomonas putida | |
| 5.3.3.1 | 4.3 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54L | Pseudomonas putida | |
| 5.3.3.1 | 7.5 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82L | Pseudomonas putida | |
| 5.3.3.1 | 11.1 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, wild-type enzyme | Pseudomonas putida | |
| 5.3.3.1 | 311 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme W116A | Pseudomonas putida | |
| 5.3.3.1 | 539 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82A | Pseudomonas putida | |
| 5.3.3.1 | 2540 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82L | Pseudomonas putida | |
| 5.3.3.1 | 3630 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54A | Pseudomonas putida | |
| 5.3.3.1 | 4420 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme W116Y | Pseudomonas putida | |
| 5.3.3.1 | 4760 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme W116F | Pseudomonas putida | |
| 5.3.3.1 | 8900 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54L | Pseudomonas putida | |
| 5.3.3.1 | 21200 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, wild-type enzyme | Pseudomonas putida | |
| 5.3.3.1 | 21230 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, wild-type enzyme | Pseudomonas putida | |
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