Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Mycobacterium tuberculosis |
Crystallization (Comment) | Organism |
---|---|
component KshA, sitting drop vapor diffusion method, using 1.1 M sodium malonate, 0.1 M HEPES, pH 7.0, and 0.5% (v/v) Jeffamine M600 | Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.024 | - |
4-androstene-3,17-dione | in 0.1 M potassium phosphate, pH 7.0, at 25°C | Mycobacterium tuberculosis | |
0.11 | - |
1,4-androstadiene-3,17-dione | in 0.1 M potassium phosphate, pH 7.0, at 25°C | Mycobacterium tuberculosis | |
1.2 | - |
O2 | in the presence of 1,4-androstadiene-3,17-dione, Km above 1.2 mM, in 0.1 M potassium phosphate, pH 7.0, at 25°C | Mycobacterium tuberculosis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P71875 | - |
- |
Mycobacterium tuberculosis H37Rv | P71875 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni2+-NTA agarose column chromatography and Source15Q column chromatography | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-androstene-3,17-dione + O2 + NADH + H+ | - |
Mycobacterium tuberculosis | 9alpha-hydroxy-4-androstene-3,17-dione + H2O + NAD+ | - |
? | |
4-androstene-3,17-dione + O2 + NADH + H+ | - |
Mycobacterium tuberculosis H37Rv | 9alpha-hydroxy-4-androstene-3,17-dione + H2O + NAD+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
3-ketosteroid 9alpha-hydroxylase | - |
Mycobacterium tuberculosis |
KshAB | - |
Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.07 | - |
4-androstene-3,17-dione | in 0.1 M potassium phosphate, pH 7.0, at 25°C | Mycobacterium tuberculosis | |
0.8 | - |
1,4-androstadiene-3,17-dione | in 0.1 M potassium phosphate, pH 7.0, at 25°C | Mycobacterium tuberculosis | |
7 | - |
O2 | in the presence of 1,4-androstadiene-3,17-dione, kcat above 7 1/sec, in 0.1 M potassium phosphate, pH 7.0, at 25°C | Mycobacterium tuberculosis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
[2Fe-2S]-center | 8.2 mol of iron and 2.4 mol of sulfur per mol of KshA protomer | Mycobacterium tuberculosis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.45 | - |
O2 | in the presence of 1,4-androstadiene-3,17-dione, in 0.1 M potassium phosphate, pH 7.0, at 25°C | Mycobacterium tuberculosis | |
3 | - |
4-androstene-3,17-dione | in 0.1 M potassium phosphate, pH 7.0, at 25°C | Mycobacterium tuberculosis | |
7.6 | - |
1,4-androstadiene-3,17-dione | in 0.1 M potassium phosphate, pH 7.0, at 25°C | Mycobacterium tuberculosis |