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Literature summary extracted from
- Multiplicity of 3-ketosteroid-9alpha-hydroxylase enzymes in Rhodococcus rhodochrous DSM43269 for specific degradation of different classes of steroids (2011), J. Bacteriol., 193, 3931-3940.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.14.15.30 | expressed in Escherichia coli BL21(DE3) and C41(DE3) cells | Rhodococcus rhodochrous |
| 1.14.15.30 | gene kshA1, encoding an isoform of the oxygenase component of 3-ketosteroid 9alpha-hydroxylase, phylogenetic analysis and tree. Transcriptional analysis and functional complementation of kshA null mutant strain RG32 by genes kshA1-55, overview. recombinant expression of kshA in Escherichia coli strains Bl21(DE3) and C41(DE3) | Rhodococcus rhodochrous |
| 1.14.15.30 | gene kshA5, encoding an isoform of the oxygenase component of 3-ketosteroid 9alpha-hydroxylase, phylogenetic analysis and tree. Transcriptional analysis and functional complementation of kshA null mutant strain RG32 by genes kshA1-55, overview. kshA5 supports growth on any of the different classes of steroids tested, consistent with its broad expression induction pattern. Recombinant expression of kshA in Escherichia coli strains Bl21(DE3) and C41(DE3) | Rhodococcus rhodochrous |
| 1.14.15.30 | gene kshB, recombinant expression of kshB in Escherichia coli strains Bl21(DE3) and C41(DE3) | Rhodococcus rhodochrous |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.14.15.30 | additional information | a 5fold kshA null mutant of strain DSM43269 (strain RG32) lacks genes kshA1, kshA2, kshA3, kshA4, and kshA5. Contrary to wild type DSM43269, the kshA null mutant strain RG32 shows no growth on progesterone, 4-androstene-3,17-dione, or cholic acid. Growth of strain RG32 on cholesterol is severely impaired, but not completely blocked | Rhodococcus rhodochrous |
| 1.14.15.30 | additional information | a 5fold kshA null mutant of strain DSM43269 (strain RG32) lacks genes kshA1, kshA2, kshA3, kshA4, and kshA5. Contrary to wild type DSM43269, the kshA null mutant strain RG32 shows no growth on progesterone, 4-androstene-3,17-dione, or cholic acid. Growth of strain RG32 on cholesterol was severely impaired, but not completely blocked | Rhodococcus rhodochrous |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.15.30 | additional information | Rhodococcus rhodochrous | all KSH enzymes with KshADSM43269 isozymes are C-9alpha-hydroxylases acting on a wide range of 3-oxosteroids, but not on 3-hydroxysteroids | ? | - |
? |  |
| 1.14.15.30 | additional information | Rhodococcus rhodochrous DSM 43269 | all KSH enzymes with KshADSM43269 isozymes are C-9alpha-hydroxylases acting on a wide range of 3-oxosteroids, but not on 3-hydroxysteroids | ? | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.15.30 | Rhodococcus rhodochrous | B6V6V5 | isozyme KshA4 | - |
| 1.14.15.30 | Rhodococcus rhodochrous | F1CMX0 | gene kshA1 encodes an isoform of the oxygenase component of 3-ketosteroid 9alpha-hydroxylase | - |
| 1.14.15.30 | Rhodococcus rhodochrous | F1CMX0 | isozyme KshA1 | - |
| 1.14.15.30 | Rhodococcus rhodochrous | F1CMX3 | gene kshB, encoding the ferredoxin reductase | - |
| 1.14.15.30 | Rhodococcus rhodochrous | F1CMX6 | isozyme KshA2 | - |
| 1.14.15.30 | Rhodococcus rhodochrous | F1CMX8 | isozyme KshA3 | - |
| 1.14.15.30 | Rhodococcus rhodochrous | F1CMY8 | gene kshA5 encodes an isoform of the oxygenase component of 3-ketosteroid 9alpha-hydroxylase | - |
| 1.14.15.30 | Rhodococcus rhodochrous | F1CMY8 | isozyme KshA5 | - |
| 1.14.15.30 | Rhodococcus rhodochrous DSM 43269 | B6V6V5 | isozyme KshA4 | - |
| 1.14.15.30 | Rhodococcus rhodochrous DSM 43269 | F1CMX0 | gene kshA1 encodes an isoform of the oxygenase component of 3-ketosteroid 9alpha-hydroxylase | - |
| 1.14.15.30 | Rhodococcus rhodochrous DSM 43269 | F1CMX0 | isozyme KshA1 | - |
| 1.14.15.30 | Rhodococcus rhodochrous DSM 43269 | F1CMX3 | gene kshB, encoding the ferredoxin reductase | - |
| 1.14.15.30 | Rhodococcus rhodochrous DSM 43269 | F1CMX6 | isozyme KshA2 | - |
| 1.14.15.30 | Rhodococcus rhodochrous DSM 43269 | F1CMX8 | isozyme KshA3 | - |
| 1.14.15.30 | Rhodococcus rhodochrous DSM 43269 | F1CMY8 | gene kshA5 encodes an isoform of the oxygenase component of 3-ketosteroid 9alpha-hydroxylase | - |
| 1.14.15.30 | Rhodococcus rhodochrous DSM 43269 | F1CMY8 | isozyme KshA5 | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.15.30 | - |
Rhodococcus rhodochrous |
| 1.14.15.30 | recombinant KshA from Escherichia coli strains Bl21(DE3) and C41(DE3) | Rhodococcus rhodochrous |
| 1.14.15.30 | recombinant KshB from Escherichia coli strains Bl21(DE3) and C41(DE3) | Rhodococcus rhodochrous |
Source Tissue
| EC Number | Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|---|
| 1.14.15.30 | additional information | cell growth on different steroids as a sole carbon and energy source with different isozymes, overview | Rhodococcus rhodochrous | - |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.15.30 | 1-(5alpha)-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 23% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-5alpha-androst-1-en-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 1-(5alpha)-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA3 shows 28% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-5alpha-androst-1-en-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 1-(5alpha)-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 68% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-5alpha-androst-1-en-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 1-(5alpha)-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA5 shows 93% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-5alpha-androst-1-en-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 1-(5alpha)-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 68% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-5alpha-androst-1-en-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 1-(5alpha)-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 23% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-5alpha-androst-1-en-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 1-(5alpha)-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA3 shows 28% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-5alpha-androst-1-en-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 1-(5alpha)-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA5 shows 93% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-5alpha-androst-1-en-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 11beta-hydrocortisone + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | i.e. stanolon, isozyme KshA5 shows 93% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-11beta-hydrocortisone + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 11beta-hydrocortisone + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | i.e. stanolon, isozyme KshA5 shows 93% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-11beta-hydrocortisone + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 19-nor-4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 24% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-19-nor-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 19-nor-4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA3 shows 10% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-19-nor-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 19-nor-4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 78% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-19-nor-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 19-nor-4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA5 shows 107% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-19-nor-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 19-nor-4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 78% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-19-nor-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 19-nor-4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 24% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-19-nor-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 19-nor-4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA3 shows 10% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-19-nor-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 19-nor-4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA5 shows 107% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-19-nor-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 500% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA3 shows 124% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 23% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA5 shows 80% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 23% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 500% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA3 shows 124% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholesta-1,4-dien-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 423% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-23,24-bisnorcholesta-1,4-dien-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholesta-1,4-dien-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA3 shows 176% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-23,24-bisnorcholesta-1,4-dien-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholesta-1,4-dien-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 22% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-23,24-bisnorcholesta-1,4-dien-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholesta-1,4-dien-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA5 shows 43% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-23,24-bisnorcholesta-1,4-dien-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholesta-1,4-dien-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 22% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-23,24-bisnorcholesta-1,4-dien-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 23,24-bisnorcholesta-1,4-dien-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA3 shows 176% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxy-23,24-bisnorcholesta-1,4-dien-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 3-oxosteroid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Rhodococcus rhodochrous | 9alpha-hydroxysteroid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 3-oxosteroid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Rhodococcus rhodochrous DSM 43269 | 9alpha-hydroxysteroid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 4-androstene-17beta-ol-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 127% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha,17beta-dihydroxy-4-androstene-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 4-androstene-17beta-ol-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA3 shows 94% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha,17beta-dihydroxy-4-androstene-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 4-androstene-17beta-ol-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 101% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha,17beta-dihydroxy-4-androstene-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 4-androstene-17beta-ol-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA5 shows 113% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha,17beta-dihydroxy-4-androstene-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 4-androstene-3,17-dione + O2 + NADH + H+ | isozyme KshA1 shows 100% activity | Rhodococcus rhodochrous | ? + H2O + NAD+ | - |
? | |
| 1.14.15.30 | 4-androstene-3,17-dione + O2 + NADH + H+ | isozyme KshA3 shows 100% activity | Rhodococcus rhodochrous | ? + H2O + NAD+ | - |
? | |
| 1.14.15.30 | 4-androstene-3,17-dione + O2 + NADH + H+ | isozyme KshA4 shows 100% activity | Rhodococcus rhodochrous | ? + H2O + NAD+ | - |
? | |
| 1.14.15.30 | 4-androstene-3,17-dione + O2 + NADH + H+ | isozyme KshA5 shows 100% activity | Rhodococcus rhodochrous | ? + H2O + NAD+ | - |
? | |
| 1.14.15.30 | 4-cholestene-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 13% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxycholest-4-en-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 4-cholestene-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA3 shows 23% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxycholest-4-en-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 4-cholestene-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA5 shows 26% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxycholest-4-en-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 4-pregnene-3,20-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 423% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-4-pregnene-3,20-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 4-pregnene-3,20-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA3 shows 112% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-4-pregnene-3,20-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 4-pregnene-3,20-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 99% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-4-pregnene-3,20-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 4-pregnene-3,20-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA5 shows 67% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-4-pregnene-3,20-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 5alpha-androstane-17beta-ol-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | i.e. stanolon, isozyme KshA5 shows 93% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha,17beta-dihydroxy-5alpha-androstane-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 5alpha-androstane-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 9% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-5alpha-androstane-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 5alpha-androstane-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 64% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-5alpha-androstane-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 5alpha-androstane-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA5 shows 80% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-5alpha-androstane-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 5beta-androstane-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 19% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-5beta-androstane-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 5beta-androstane-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 58% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-5beta-androstane-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 5beta-androstane-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA5 shows 93% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxy-5beta-androstane-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 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 | - |
? | |
| 1.14.15.30 | androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA1 shows 288% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA3 shows 194% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA4 shows 89% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | isozyme KshA5 shows 49% activity compared to 4-androstene-3,17-dione | Rhodococcus rhodochrous | 9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
| 1.14.15.30 | 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 | - |
? | |
| 1.14.15.30 | additional information | isozyme KshA5 is the most versatile isoenzyme, with the broadest substrate range but without a clear substrate preference. It shows no activity with 5-cholestene-3beta-ol (cholesterol), 5alpha-androstane-17-one, 3alpha-hydroxy-5alpha-pregnane-20-one, and 3beta-hydroxy-5alpha-androstane-17-one | Rhodococcus rhodochrous | ? | - |
? | |
| 1.14.15.30 | additional information | no activity with 4-cholestene-3-one, 5-cholestene-3beta-ol (cholesterol), 5alpha-androstane-17-one, 3alpha-hydroxy-5alpha-pregnane-20-one, 3beta-hydroxy-5alpha-androstane-17-one, 5alpha-androstane-17beta-ol-3-one (stanolon) and 11beta-hydrocortisone | Rhodococcus rhodochrous | ? | - |
? | |
| 1.14.15.30 | additional information | no activity with 5-cholestene-3beta-ol (cholesterol), 5alpha-androstane-17-one, 3alpha-hydroxy-5alpha-pregnane-20-one, 3beta-hydroxy-5alpha-androstane-17-one, 5alpha-androstane-17beta-ol-3-one (stanolon) and 11beta-hydrocortisone | Rhodococcus rhodochrous | ? | - |
? | |
| 1.14.15.30 | additional information | no activity with 5alpha-androstane-3,17-dione, 5beta-androstane-3,17-dione, 5-cholestene-3beta-ol (cholesterol), 5alpha-androstane-17-one, 3alpha-hydroxy-5alpha-pregnane-20-one, 3beta-hydroxy-5alpha-androstane-17-one, 5alpha-androstane-17beta-ol-3-one (stanolon) and 11beta-hydrocortisone | Rhodococcus rhodochrous | ? | - |
? | |
| 1.14.15.30 | additional information | all KSH enzymes with KshADSM43269 isozymes are C-9alpha-hydroxylases acting on a wide range of 3-oxosteroids, but not on 3-hydroxysteroids | Rhodococcus rhodochrous | ? | - |
? | |
| 1.14.15.30 | additional information | no activity with 4-cholestene-3-one, 5-cholestene-3beta-ol (cholesterol), 5alpha-androstane-17-one, 3alpha-hydroxy-5alpha-pregnane-20-one, 3beta-hydroxy-5alpha-androstane-17-one, 5alpha-androstane-17beta-ol-3-one (stanolon) and 11beta-hydrocortisone | Rhodococcus rhodochrous DSM 43269 | ? | - |
? | |
| 1.14.15.30 | additional information | no activity with 5-cholestene-3beta-ol (cholesterol), 5alpha-androstane-17-one, 3alpha-hydroxy-5alpha-pregnane-20-one, 3beta-hydroxy-5alpha-androstane-17-one, 5alpha-androstane-17beta-ol-3-one (stanolon) and 11beta-hydrocortisone | Rhodococcus rhodochrous DSM 43269 | ? | - |
? | |
| 1.14.15.30 | additional information | all KSH enzymes with KshADSM43269 isozymes are C-9alpha-hydroxylases acting on a wide range of 3-oxosteroids, but not on 3-hydroxysteroids | Rhodococcus rhodochrous DSM 43269 | ? | - |
? | |
| 1.14.15.30 | additional information | no activity with 5alpha-androstane-3,17-dione, 5beta-androstane-3,17-dione, 5-cholestene-3beta-ol (cholesterol), 5alpha-androstane-17-one, 3alpha-hydroxy-5alpha-pregnane-20-one, 3beta-hydroxy-5alpha-androstane-17-one, 5alpha-androstane-17beta-ol-3-one (stanolon) and 11beta-hydrocortisone | Rhodococcus rhodochrous DSM 43269 | ? | - |
? | |
| 1.14.15.30 | additional information | isozyme KshA5 is the most versatile isoenzyme, with the broadest substrate range but without a clear substrate preference. It shows no activity with 5-cholestene-3beta-ol (cholesterol), 5alpha-androstane-17-one, 3alpha-hydroxy-5alpha-pregnane-20-one, and 3beta-hydroxy-5alpha-androstane-17-one | Rhodococcus rhodochrous DSM 43269 | ? | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.14.15.30 | 3-ketosteroid 9alpha-hydroxylase | - |
Rhodococcus rhodochrous |
| 1.14.15.30 | 3-ketosteroid-9alpha-hydroxylase | - |
Rhodococcus rhodochrous |
| 1.14.15.30 | KSH | - |
Rhodococcus rhodochrous |
| 1.14.15.30 | KshA1 | isozyme | Rhodococcus rhodochrous |
| 1.14.15.30 | KshA2 | isoenzyme | Rhodococcus rhodochrous |
| 1.14.15.30 | KshA3 | isoenzyme | Rhodococcus rhodochrous |
| 1.14.15.30 | KshA4 | isoenzyme | Rhodococcus rhodochrous |
| 1.14.15.30 | KshA5 | isoenzyme | Rhodococcus rhodochrous |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.15.30 | 7 | - |
assay at | Rhodococcus rhodochrous |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.15.30 | [2Fe-2S]-center | - |
Rhodococcus rhodochrous | |
Expression
| EC Number | Organism | Comment | Expression |
|---|---|---|---|
| 1.14.15.30 | Rhodococcus rhodochrous | in wild type Rhodococcus rhodochrous strain DSM43269, ksh1 is specifically induced by cholic acid | up |
| 1.14.15.30 | Rhodococcus rhodochrous | in wild type Rhodococcus rhodochrous strain DSM43269, kshA2 is specifically induced by 4-androstene-3,17-dione, progesterone, and cholic acid | up |
| 1.14.15.30 | Rhodococcus rhodochrous | in wild type Rhodococcus rhodochrous strain DSM43269, kshA3 is specifically induced by 4-androstene-3,17-dione | up |
| 1.14.15.30 | Rhodococcus rhodochrous | in wild type Rhodococcus rhodochrous strain DSM43269, kshA4 is specifically induced by 4-androstene-3,17-dione, cholic acid, and cholesterol | up |
| 1.14.15.30 | Rhodococcus rhodochrous | in wild type Rhodococcus rhodochrous strain DSM43269, kshA5 is specifically induced by 4-androstene-3,17-dione, progesterone, cholic acid, and cholesterol | up |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 1.14.15.30 | metabolism | 3-ketosteroid-9alpha-hydroxylase is a key enzyme in steroid catabolism | Rhodococcus rhodochrous |
| 1.14.15.30 | additional information | KshA and KshB together constitute the KSH enzyme | Rhodococcus rhodochrous |
| 1.14.15.30 | physiological function | 3-ketosteroid 9alpha-hydroxylase is a key enzyme in steroid catabolism | Rhodococcus rhodochrous |
| 1.14.15.30 | physiological function | 3-ketosteroid 9alpha-hydroxylase is a key enzyme in steroid catabolism. Isozyme KshA1 is dedicated to cholic acid catabolism. Presence of multiple kshA genes in the Rhodococcus rhodochrous DSM43269 genome, each displaying unique steroid induction patterns and substrate ranges, appears to facilitate a dynamic and fine-tuned steroid catabolism, with C-9 alpha-hydroxylation occurring at different levels during microbial steroid degradation | Rhodococcus rhodochrous |
| 1.14.15.30 | physiological function | 3-ketosteroid 9alpha-hydroxylase is a key enzyme in steroid catabolism. Isozyme KshA5 appears to be the most versatile enzyme, with the broadest substrate range but without a clear substrate preference. Presence of multiple kshA genes in the Rhodococcus rhodochrous DSM 43269 genome, each displaying unique steroid induction patterns and substrate ranges, appears to facilitate a dynamic and fine-tuned steroid catabolism, with C-9 alpha-hydroxylation occurring at different levels during microbial steroid degradation | Rhodococcus rhodochrous |
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Release 2023.1 (January 2023)
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