Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli GJ1158 cells | Rhodococcus rhodochrous |
Crystallization (Comment) | Organism |
---|---|
in complex with 1,4-androstadiene-3,17-dione or 3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA, hanging drop vapor diffusion method, using 500 mM NaH2PO4, 125 mM K2HPO4, 4% (w/v) PEG-1000 or 2% (w/v) PEG-3000, 20 mM Tris, 100 mM phosphate-citrate, pH 4.2 | Rhodococcus rhodochrous |
purified recombinant enzyme in complex with substrate androsta-1,4-diene-3,17-dione, using 500 mM NaH2PO4, 125 mM K2HPO4, 4% PEG 1000, 20 mM Tris, 100 mM phosphate-citrate, pH 4.2, X-ray diffraction structure determination and analysis at 2.46-2.52 A resolution | Mycobacterium tuberculosis |
purified recombinant enzyme in complex with substrates androsta-1,4-diene-3,17-dione and 3-oxo-23,24-bisnorcholesta-1,4-diene-22-oate and enzyme KshA5. Attempts to crystallize KshA1 alone are unsuccessful, crystals are serendipitously obtained from a mixture of KshA1, KshA5, and 1,4-BNC-CoA. Pyramid-shaped light brown crystals grow over 2 weeks from a 0.0015 ml drop containing equal volumes of KshA1 with 1,4-BNC-CoA, KshA5, and well solution containing 500 mM NaH2PO4, 125 mM K2HPO4, 4% PEG 1000, 20 mM Tris, 100 mM phosphate-citrate, pH 4.2, X-ray diffraction structure determination and analysis at 2.45-2.52 A resolution | Rhodococcus rhodochrous |
purified recombinant enzyme in complex with substrates androsta-1,4-diene-3,17-dione and 3-oxo-23,24-bisnorcholesta-1,4-diene-22-oate, KshA5 is crystallized in complex with 1,4-BNC-CoA using 0.8 M NaH2PO4, 0.2 M K2HPO4, 0.25 M (NH4)2SO4, 20 mM CHES, 40 mM NaCl, and 0.1 M phosphate-citrate, pH 4.2, and crystallized in complex with ADD using 0.8 M NaH2PO4, 0.2 M K2HPO4, 2% PEG 3000, 20 mM CHES, 0.1 M phosphate-citrate, pH 4.2, X-ray diffraction structure determination and analysis at 1.92 A and 2.6 A resolution, respectively | Rhodococcus rhodochrous |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
3-oxo-23,24-bisnorchol-4-en-22-oate | substrate inhibition | Rhodococcus rhodochrous | |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | strong substrate inhibition; strong substrate inhibition | Rhodococcus rhodochrous | |
4-androstene-3,17-dione | strong substrate inhibition; strong substrate inhibition | Rhodococcus rhodochrous | |
testosterone | substrate inhibition | Rhodococcus rhodochrous |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetic analysis | Rhodococcus rhodochrous | |
additional information | - |
additional information | steady-state kinetic analysis | Mycobacterium tuberculosis | |
0.0005 | - |
testosterone | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.0005 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.0005 | - |
testosterone | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.0005 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.0008 | - |
4-Estren-3,17-dione | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.0008 | - |
4-Estren-3,17-dione | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.001 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.001 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.0012 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.0012 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.0015 | - |
O2 | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.0015 | - |
O2 | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.002 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.002 | - |
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.0022 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.0022 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.0061 | - |
5alpha-androstan-3,17-dione | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.049 | - |
O2 | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.049 | - |
O2 | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.05 | - |
androsta-1,4-diene-3,17-dione | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.05 | - |
1,4-androstadiene-3,17-dione | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.11 | - |
testosterone | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.11 | - |
testosterone | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.4 | - |
4-Estren-3,17-dione | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.4 | - |
4-Estren-3,17-dione | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.5 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.5 | - |
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | Rhodococcus rhodochrous | - |
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | Mycobacterium tuberculosis | - |
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | Mycobacterium tuberculosis H37Rv | - |
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | Rhodococcus rhodochrous DSM 43269 | - |
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P71875 | KshA | - |
Mycobacterium tuberculosis H37Rv | P71875 | KshA | - |
Rhodococcus rhodochrous | F1CMX0 | - |
- |
Rhodococcus rhodochrous | F1CMX0 | KshA1 | - |
Rhodococcus rhodochrous | F1CMY8 | - |
- |
Rhodococcus rhodochrous | F1CMY8 | KshA5 | - |
Rhodococcus rhodochrous DSM 43269 | F1CMX0 | - |
- |
Rhodococcus rhodochrous DSM 43269 | F1CMX0 | KshA1 | - |
Rhodococcus rhodochrous DSM 43269 | F1CMY8 | - |
- |
Rhodococcus rhodochrous DSM 43269 | F1CMY8 | KshA5 | - |
Purification (Comment) | Organism |
---|---|
Ni-Sepharose column chromatography and Source 15Q column chromatography | Rhodococcus rhodochrous |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 = 9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | KshAB-catalyzed 9alpha-hydroxylation of 3-ketosteroids. 1,4-Diene ketosteroids undergo subsequent nonenzymatic cleavage of ring B and aromatization of ring A | Rhodococcus rhodochrous | |
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 = 9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | KshAB-catalyzed 9alpha-hydroxylation of 3-ketosteroids. 1,4-Diene ketosteroids undergo subsequent nonenzymatic cleavage of ring B and aromatization of ring A | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,4-androstadiene-3,17-dione + reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Rhodococcus rhodochrous | 3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
1,4-androstadiene-3,17-dione + reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Rhodococcus rhodochrous DSM 43269 | 3-hydroxy-9,10-seconandrost-1,3,5(10)-triene-9,17-dione + oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
3-oxo-23,24-bisnorchol-4-en-22-oate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Rhodococcus rhodochrous | 9alpha-hydroxy-3-oxo-23,24-bisnorchol-4-en-22-oate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
3-oxo-23,24-bisnorchol-4-en-22-oate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-3-oxo-23,24-bisnorchol-4-en-22-oate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Rhodococcus rhodochrous | ? | - |
? | |
3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Rhodococcus rhodochrous DSM 43269 | ? | - |
? | |
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA + reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Rhodococcus rhodochrous | ? | - |
? | |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | low activity | Rhodococcus rhodochrous | 9alpha-hydroxy-3-oxo-23,24-bisnorchola-1,4-diene-22-oate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Rhodococcus rhodochrous | ? | - |
? | |
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Rhodococcus rhodochrous | 9alpha-hydroxy-3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
4-androstene-3,17-dione + reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Rhodococcus rhodochrous | 9alpha-hydroxy-4-androstene-3,17-dione + oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
4-estren-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Rhodococcus rhodochrous | 9alpha-hydroxy-4-estren-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
4-estren-3,17-dione + reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Rhodococcus rhodochrous | 9alpha-hydroxy-4-estren-3,17-dione + oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
4-estren-3,17-dione + reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Rhodococcus rhodochrous | 9alpha-hydroxy-4-estren-3,17-dione + oxidized ferredoxin [iron-sulfur] cluster + H2O | ? | ? | |
5alpha-androstan-3,17-dione + reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Rhodococcus rhodochrous | 9alpha-hydroxy-5alpha-androstane-3,17-dione + oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Rhodococcus rhodochrous | 9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Mycobacterium tuberculosis | 9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Mycobacterium tuberculosis H37Rv | 9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
additional information | cholesterol-degrading KshAMtb from Mycobacterium tuberculosis has the highest catalytic efficiency for CoA-thioesterified substrates, substrate specificity comparisons of KSH enzymes, overview | Mycobacterium tuberculosis | ? | - |
? | |
additional information | enzyme KshA1 has the highest apparent catalytic efficiency (kcat/Km) for steroids with an isopropyl side chain at C17, such as 3-oxo-23,24-bisnorcholesta-1,4-diene-22-oate, substrate specificity comparisons of KSH enzymes, overview | Rhodococcus rhodochrous | ? | - |
? | |
additional information | enzyme KshA5 has the highest apparent catalytic efficiency for substrates with no C17 side chain, e.g. 4-estrendione, 5alpha-androstandione, and testosterone, substrate specificity comparisons of KSH enzymes, overview | Rhodococcus rhodochrous | ? | - |
? | |
additional information | cholesterol-degrading KshAMtb from Mycobacterium tuberculosis has the highest catalytic efficiency for CoA-thioesterified substrates, substrate specificity comparisons of KSH enzymes, overview | Mycobacterium tuberculosis H37Rv | ? | - |
? | |
additional information | enzyme KshA1 has the highest apparent catalytic efficiency (kcat/Km) for steroids with an isopropyl side chain at C17, such as 3-oxo-23,24-bisnorcholesta-1,4-diene-22-oate, substrate specificity comparisons of KSH enzymes, overview | Rhodococcus rhodochrous DSM 43269 | ? | - |
? | |
additional information | enzyme KshA5 has the highest apparent catalytic efficiency for substrates with no C17 side chain, e.g. 4-estrendione, 5alpha-androstandione, and testosterone, substrate specificity comparisons of KSH enzymes, overview | Rhodococcus rhodochrous DSM 43269 | ? | - |
? | |
testosterone + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Rhodococcus rhodochrous | 9alpha,17beta-dihydroxy-4-androstene-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
testosterone + reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Rhodococcus rhodochrous | (17beta)-9,17-dihydroxyandrost-4-en-3-one + oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
testosterone + reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Rhodococcus rhodochrous | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | KshAB is a two-component Rieske oxygenase comprising an oxygenase (KshA) and a reductase (KshB), structures comparisons of different KSH enzymes | Rhodococcus rhodochrous |
More | KshAB is a two-component Rieske oxygenase comprising an oxygenase (KshA) and a reductase (KshB), structures comparisons of different KSH enzymes. The substrate binding residues within the active site pocket are positioned nearly identically in enzyme KshA1 from Rhodococcus rhodochrous and KshA from Mycobacterium tuberculosis | Rhodococcus rhodochrous |
More | KshAB is a two-component Rieske oxygenase comprising an oxygenase (KshA) and a reductase (KshB), structures comparisons of different KSH enzymes. The substrate binding residues within the active site pocket are positioned nearly identically in enzyme KshA1 from Rhodococcus rhodochrous and KshA from Mycobacterium tuberculosis | Mycobacterium tuberculosis |
Synonyms | Comment | Organism |
---|---|---|
3-ketosteroid 9alpha-hydroxylase | - |
Rhodococcus rhodochrous |
3-ketosteroid 9alpha-hydroxylase | - |
Mycobacterium tuberculosis |
KshA | - |
Mycobacterium tuberculosis |
KshA1 | - |
Rhodococcus rhodochrous |
KshA1 | oxygenase component of 3-ketosteroid 9alpha-hydroxylase | Rhodococcus rhodochrous |
KshA5 | - |
Rhodococcus rhodochrous |
KshA5 | oxygenase component of 3-ketosteroid 9alpha-hydroxylase | Rhodococcus rhodochrous |
KshAB | - |
Rhodococcus rhodochrous |
KshAB | - |
Mycobacterium tuberculosis |
KshAMt | - |
Mycobacterium tuberculosis |
Rv3526 | - |
Mycobacterium tuberculosis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at | Rhodococcus rhodochrous |
22 | - |
assay at | Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.5 | - |
4-Estren-3,17-dione | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.5 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.5 | - |
4-Estren-3,17-dione | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.5 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.6 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.6 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.7 | - |
androsta-1,4-diene-3,17-dione | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.7 | - |
1,4-androstadiene-3,17-dione | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.8 | - |
O2 | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.8 | - |
testosterone | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.8 | - |
4-Estren-3,17-dione | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.8 | - |
O2 | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.8 | - |
testosterone | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.8 | - |
5alpha-androstan-3,17-dione | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.8 | - |
4-Estren-3,17-dione | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
0.9 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.9 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
1.1 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA | pH 7.0, 22°C | Rhodococcus rhodochrous | |
1.1 | - |
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
1.3 | - |
androsta-1,4-diene-3,17-dione | pH 7.0, 22°C | Rhodococcus rhodochrous | |
1.3 | - |
1,4-androstadiene-3,17-dione | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
1.4 | - |
testosterone | pH 7.0, 22°C | Rhodococcus rhodochrous | |
1.4 | - |
testosterone | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
1.7 | - |
O2 | pH 7.0, 22°C | Rhodococcus rhodochrous | |
1.7 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA | pH 7.0, 22°C | Rhodococcus rhodochrous | |
1.7 | - |
O2 | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
1.7 | - |
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
2.6 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
2.6 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Rhodococcus rhodochrous |
7 | - |
assay at | Mycobacterium tuberculosis |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.11 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
0.13 | - |
testosterone | pH 7.0, 22°C | Rhodococcus rhodochrous |
General Information | Comment | Organism |
---|---|---|
additional information | the enzyme has a steroid-binding pocket with the substrate's C17 oriented toward the active site opening. Comparisons suggest that Tyr245 and Phe297 are determinants of KshA1 specificity. The enzyme has a flexible 16-residue mouth loop, which in some structures completely occlude the substrate-binding pocket from the bulk solvent | Rhodococcus rhodochrous |
additional information | the enzyme has a steroid-binding pocket with the substrate's C17 oriented toward the active site opening. Comparisons suggest that Tyr245 and Phe297 are determinants of KshA1 specificity. The enzyme has a flexible 16-residue mouth loop, which in some structures completely occlude the substrate-binding pocket from the bulk solvent | Mycobacterium tuberculosis |
additional information | the enzyme has a steroid-binding pocket with the substrate's C17 oriented toward the active site opening. The enzyme has a flexible 16-residue mouth loop, which in some structures completely occlude the substrate-binding pocket from the bulk solvent. The catalytic iron andalpha-helices harboring its ligands are displaced up to 4.4 A in theKshA5substrate complexes as compared with substrate-free KshA, suggesting that Rieske oxygenases may have a dynamic nature similar to cytochrome P450 | Rhodococcus rhodochrous |
physiological function | KshA is the oxygenase component of 3-ketosteroid 9alpha-hydroxylase, a Rieske oxygenase involved in the bacterial degradation of steroids | Mycobacterium tuberculosis |
physiological function | KshA is the oxygenase component of 3-ketosteroid 9alpha-hydroxylase, a Rieske oxygenase involved in the bacterial degradation of steroids | Rhodococcus rhodochrous |
physiological function | KshA is the oxygenase component of 3-ketosteroid 9alpha-hydroxylase, a Rieske oxygenase involved in the bacterial degradation of steroids. KshA1 has a role in bile acid catabolism | Rhodococcus rhodochrous |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.2 | - |
4-Estren-3,17-dione | pH 7.0, 22°C | Rhodococcus rhodochrous | |
1.2 | - |
4-Estren-3,17-dione | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
3.7 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA | pH 7.0, 22°C | Rhodococcus rhodochrous | |
3.7 | - |
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
13 | - |
testosterone | pH 7.0, 22°C | Rhodococcus rhodochrous | |
13 | - |
testosterone | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
17 | - |
O2 | pH 7.0, 22°C | Rhodococcus rhodochrous | |
17 | - |
O2 | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
27 | - |
androsta-1,4-diene-3,17-dione | pH 7.0, 22°C | Rhodococcus rhodochrous | |
27 | - |
1,4-androstadiene-3,17-dione | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
140 | - |
5alpha-androstan-3,17-dione | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
500 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
500 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
600 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA | pH 7.0, 22°C | Rhodococcus rhodochrous | |
600 | - |
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
900 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
900 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
1000 | - |
4-Estren-3,17-dione | pH 7.0, 22°C | Rhodococcus rhodochrous | |
1000 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
1000 | - |
4-Estren-3,17-dione | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
1000 | - |
3-oxo-23,24-bisnorchola-1,4-diene-22-oate | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
1100 | - |
O2 | pH 7.0, 22°C | Rhodococcus rhodochrous | |
1100 | - |
O2 | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
1200 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | pH 7.0, 22°C | Rhodococcus rhodochrous | |
1200 | - |
3-oxo-23,24-bisnorchol-4-en-22-oate | isoform KshA1, at pH 7.0 and 22°C | Rhodococcus rhodochrous | |
1500 | - |
testosterone | pH 7.0, 22°C | Rhodococcus rhodochrous | |
1500 | - |
testosterone | isoform KshA5, at pH 7.0 and 22°C | Rhodococcus rhodochrous |