Crystallization (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
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 |
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 |
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 |
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 |
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 |
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 |
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] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0149 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82A | Pseudomonas putida | |
0.0328 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82A | Pseudomonas putida | |
0.0499 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, wild-type enzyme | Pseudomonas putida | |
0.0539 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82L | Pseudomonas putida | |
0.0579 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54A | Pseudomonas putida | |
0.0651 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82L | Pseudomonas putida | |
0.077 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54L | Pseudomonas putida | |
0.0982 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54L | Pseudomonas putida | |
0.11 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, wild-type enzyme | Pseudomonas putida | |
0.149 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme W116F | Pseudomonas putida | |
0.174 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme W116A | Pseudomonas putida | |
0.256 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54A | Pseudomonas putida |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | - |
biotype B | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5(10)-estrene-3,17-dione | - |
Pseudomonas putida | estr-4-en-3,17-dione | - |
? | |
5-Androstene-3,17-dione | - |
Pseudomonas putida | 4-Androstene-3,17-dione | - |
? | |
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 Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.25 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82A | Pseudomonas putida | |
1.7 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54A | Pseudomonas putida | |
4.3 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54L | Pseudomonas putida | |
7.5 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82L | Pseudomonas putida | |
11.1 | - |
5(10)-estrene-3,17-dione | 25°C, pH 7.0, wild-type enzyme | Pseudomonas putida | |
311 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme W116A | Pseudomonas putida | |
539 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82A | Pseudomonas putida | |
2540 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F82L | Pseudomonas putida | |
3630 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54A | Pseudomonas putida | |
4420 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme W116Y | Pseudomonas putida | |
4760 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme W116F | Pseudomonas putida | |
8900 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme F54L | Pseudomonas putida | |
21200 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, wild-type enzyme | Pseudomonas putida | |
21230 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, wild-type enzyme | Pseudomonas putida |