Crystallization (Comment) | Organism |
---|---|
crystal structure of the R72A mutant enzyme determined at 2.5 A resolution belongs to the space group C2221 with cell dimensions of a = 36.37 A, b = 74.44 A and c = 96.06 A. Crystals are grown from a solution containing 2.0 M ammonioum acetate and 0.1 M sodium acetate at pH 4.6 by hanging drop vapor-diffusion method at 22°C | Pseudomonas putida |
Protein Variants | Comment | Organism |
---|---|---|
E118A | the free-energy change for unfolding in the absence of urea at 25°C is decreased by about 3.9 kcal/mol compared to wild-type value. Mutation increasex the dissociation constant for (+)-equilenin, a reaction intermediate analogue. 50% of the protein is unfolded at 4.46 M urea compared to 5.22 M for the wild-type enzyme. The turnover-number for 5-androstene-3,17-dione is 33% of that of the wild-type enzyme, the Km-value is 348% of that of the wild-type enzyme | Pseudomonas putida |
E118A/N120A | the free-energy change for unfolding in the absence of urea at 25°C is decreased by about 9.5 kcal/mol compared to wild-type value. Mutation increases the dissociation constant for (+)-equilenin, a reaction intermediate analogue. 50% of the protein is unfolded at 3.89 M urea compared to 5.22 M for the wild-type enzyme. The turnover-number for 5-androstene-3,17-dione is 6% of that of the wild-type enzyme, the Km-value is 523% of that of the wild-type enzyme | Pseudomonas putida |
N120A | the free-energy change for unfolding in the absence of urea at 25°C is decreased by about 7.8 kcal/mol compared to wild-type value. Mutation increasex the dissociation constant for (+)-equilenin, a reaction intermediate analogue. 50% of the protein is unfolded at 3.95 M urea compared to 5.22 M for the wild-type enzyme. The turnover-number for 5-androstene-3,17-dione is 17% of that of the wild-type enzyme, the Km-value is 516% of that of the wild-type enzyme | Pseudomonas putida |
R72A | the free-energy change for unfolding in the absence of urea at 25°C is decreased by about 3.8 kcal/mol compared to wild-type value. Mutation increasex the dissociation constant for (+)-equilenin, a reaction intermediate analogue. 50% of the protein is unfolded at 4.74 M urea compared to 5.22 M for the wild-type enzyme. The turnover-number for 5-androstene-3,17-dione is 23% of that of the wild-type enzyme | Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0499 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, wild-type enzyme | Pseudomonas putida | |
0.173 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme E118A | Pseudomonas putida | |
0.258 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme N120A | Pseudomonas putida | |
0.261 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme E118A/N120A | Pseudomonas putida |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
14000 | - |
2 * 14000, mutant enzymnes R72A, E118A, N120A and E18A/N120A, SDS-PAGE | Pseudomonas putida |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | - |
biotype B | - |
Renatured (Comment) | Organism |
---|---|
50% of the wild-type enzyme is unfolded at 5.22 M urea. Mutant enzymes R72A, E118A, N120A and E118A/N120A are unfolded at lower concentrations | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5-Androstene-3,17-dione | - |
Pseudomonas putida | 4-Androstene-3,17-dione | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 14000, mutant enzymnes R72A, E118A, N120A and E18A/N120A, SDS-PAGE | Pseudomonas putida |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
the free-energy change for unfolding in the absence of urea at 25°C is 24.4 kcal/mol for the wild-type enzyme | Pseudomonas putida |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1180 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme E118A/N120A | Pseudomonas putida | |
3710 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme N120A | Pseudomonas putida | |
4870 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme R72A | Pseudomonas putida | |
6920 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, mutant enzyme E118A | Pseudomonas putida | |
21200 | - |
5-androstene-3,17-dione | pH 7.0, 25°C, wild-type enzyme | Pseudomonas putida |