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Literature summary extracted from
- Pseudoreversion of the catalytic activity of Y14F by the additional substitution(s) of tyrosine with phenylalanine in the hydrogen bond network of DELTA 5-3-ketosteroid isomerase from Pseudomonas putida biotype B (2001), Biochemistry, 40, 6828-6835.
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 5.3.3.1 | hanging drop vapor diffusion method. Crystal structures of Y14F and Y14F/Y30F/Y55F are determined at 1.8 and 2.0 A resolution, respectively | Pseudomonas putida |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 5.3.3.1 | Y14F | the hydrogen bond between Asp99 Odelta2 and C3-O of the steroid, which is perturbed by the Y14F mutation, can be partially restored to that in the wild-type enzyme by additional Y30F/Y55F mutations | Pseudomonas putida |
| 5.3.3.1 | Y14F/Y30F | the turnover-number for 5-androstene-3,17-dione is about 1% of that of the wild-type enzyme, the Km-value is 1.57fold higher than that of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | Y14F/Y30F/Y55F | the turnover-number for 5-androstene-3,17-dione is about 2.1% of that of the wild-type enzyme, the Km-value is comparable to the Km-value of the wild-type enzyme. The hydrogen bond between Asp99 Odelta2 and C3-O of the steroid, which is perturbed by the Y14F mutation, can be partially restored to that in the wild-type enzyme by additional Y30F/Y55F mutations. The improvement in the catalytic activity of Y14F by the additional Y30f/Y55F mutation is due to the changes in the structural integrity at the catalytic site and the resulting restoration of the proton-transfer mechanism in Y14F/Y30F/Y55F | Pseudomonas putida |
| 5.3.3.1 | Y14F/Y30F/Y55F/D99L | the turnover-number for 5-androstene-3,17-dione is less than 1.2% of that of the wild-type enzyme, the Km-value is 1.8fold higher than that of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | Y14F/Y30F/Y55F/D99N | the turnover-number for 5-androstene-3,17-dione is less than 1% of that of the wild-type enzyme, the Km-value is 1.14fold higher than that of the wild-type enzyme | Pseudomonas putida |
| 5.3.3.1 | Y14F/Y55F | the turnover-number for 5-androstene-3,17-dione is about 1.2% of that of the wild-type enzyme, the Km-value is 1.75fold higher than that 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 | 0.0287 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme Y14F/Y55F | Pseudomonas putida |  |
| 5.3.3.1 | 0.0503 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, wild-type enzyme | Pseudomonas putida | |
| 5.3.3.1 | 0.0513 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme Y14F/Y30F/Y55F | Pseudomonas putida | |
| 5.3.3.1 | 0.0578 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme Y14F/Y30F/Y55F/D99N | Pseudomonas putida | |
| 5.3.3.1 | 0.0787 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme Y14F/Y30F | Pseudomonas putida | |
| 5.3.3.1 | 0.0918 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme Y14F/Y30F/Y55F/D99L | 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-Androstene-3,17-dione | - |
Pseudomonas putida | 4-Androstene-3,17-dione | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 5.3.3.1 | DELTA5-ketosteroid isomerase | - |
Pseudomonas putida |
| 5.3.3.1 | KSI | - |
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 | 0.052 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme Y14F/Y30F/Y55F/D99L | Pseudomonas putida | |
| 5.3.3.1 | 1.1 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme Y14F/Y30F/Y55F/D99N | Pseudomonas putida | |
| 5.3.3.1 | 302 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme Y14F/Y30F | Pseudomonas putida | |
| 5.3.3.1 | 360 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme Y14F/Y55F | Pseudomonas putida | |
| 5.3.3.1 | 587 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, mutant enzyme Y14F/Y30F/Y55F | Pseudomonas putida | |
| 5.3.3.1 | 27900 | - |
5-androstene-3,17-dione | 25°C, pH 7.0, wild-type enzyme | Pseudomonas putida | |
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