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Literature summary extracted from

  • Penfield, J.S.; Worrall, L.J.; Strynadka, N.C.; Eltis, L.D.
    Substrate specificities and conformational flexibility of 3-ketosteroid 9alpha-hydroxylases (2014), J. Biol. Chem., 289, 25523-25536 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.15.30 expressed in Escherichia coli GJ1158 cells Rhodococcus rhodochrous

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.14.15.30 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
1.14.15.30 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
1.14.15.30 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
1.14.15.30 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

EC Number Inhibitors Comment Organism Structure
1.14.15.30 3-oxo-23,24-bisnorchol-4-en-22-oate substrate inhibition Rhodococcus rhodochrous
1.14.15.30 3-oxo-23,24-bisnorchola-1,4-diene-22-oate strong substrate inhibition; strong substrate inhibition Rhodococcus rhodochrous
1.14.15.30 4-androstene-3,17-dione strong substrate inhibition; strong substrate inhibition Rhodococcus rhodochrous
1.14.15.30 testosterone substrate inhibition Rhodococcus rhodochrous

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.15.30 additional information
-
 additional information steady-state kinetic analysis Rhodococcus rhodochrous
1.14.15.30 additional information
-
 additional information steady-state kinetic analysis Mycobacterium tuberculosis
1.14.15.30 0.0005
-
 testosterone pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.0005
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.0005
-
 testosterone isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.0005
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.0008
-
 4-Estren-3,17-dione pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.0008
-
 4-Estren-3,17-dione isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.001
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.001
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.0012
-
 3-oxo-23,24-bisnorchol-4-en-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.0012
-
 3-oxo-23,24-bisnorchol-4-en-22-oate isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.0015
-
 O2 pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.0015
-
 O2 isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.002
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.002
-
 3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.0022
-
 3-oxo-23,24-bisnorchol-4-en-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.0022
-
 3-oxo-23,24-bisnorchol-4-en-22-oate isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.0061
-
 5alpha-androstan-3,17-dione isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.049
-
 O2 pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.049
-
 O2 isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.05
-
 androsta-1,4-diene-3,17-dione pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.05
-
 1,4-androstadiene-3,17-dione isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.11
-
 testosterone pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.11
-
 testosterone isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.4
-
 4-Estren-3,17-dione pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.4
-
 4-Estren-3,17-dione isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.5
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 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/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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 Mycobacterium tuberculosis
-
 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 Mycobacterium tuberculosis H37Rv
-
 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
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.15.30 Mycobacterium tuberculosis P71875 KshA
-
1.14.15.30 Mycobacterium tuberculosis H37Rv P71875 KshA
-
1.14.15.30 Rhodococcus rhodochrous F1CMX0
-
-
1.14.15.30 Rhodococcus rhodochrous F1CMX0 KshA1
-
1.14.15.30 Rhodococcus rhodochrous F1CMY8
-
-
1.14.15.30 Rhodococcus rhodochrous F1CMY8 KshA5
-
1.14.15.30 Rhodococcus rhodochrous DSM 43269 F1CMX0
-
-
1.14.15.30 Rhodococcus rhodochrous DSM 43269 F1CMX0 KshA1
-
1.14.15.30 Rhodococcus rhodochrous DSM 43269 F1CMY8
-
-
1.14.15.30 Rhodococcus rhodochrous DSM 43269 F1CMY8 KshA5
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.15.30 Ni-Sepharose column chromatography and Source 15Q column chromatography Rhodococcus rhodochrous

Reaction

EC Number Reaction Comment Organism Reaction ID
1.14.15.30 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
a
1.14.15.30 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
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.15.30 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
-
 ?
1.14.15.30 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
-
 ?
1.14.15.30 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
-
 ?
1.14.15.30 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
-
 ?
1.14.15.30 3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
 Rhodococcus rhodochrous ?
-
 ?
1.14.15.30 3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
 Rhodococcus rhodochrous DSM 43269 ?
-
 ?
1.14.15.30 3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
 Rhodococcus rhodochrous ?
-
 ?
1.14.15.30 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
-
 ?
1.14.15.30 3-oxo-23,24-bisnorchola-1,4-diene-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
 Rhodococcus rhodochrous ?
-
 ?
1.14.15.30 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
-
 ?
1.14.15.30 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
-
 ?
1.14.15.30 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
-
 ?
1.14.15.30 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
-
 ?
1.14.15.30 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 ? ?
1.14.15.30 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
-
 ?
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
-
 Mycobacterium tuberculosis 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
-
 Mycobacterium tuberculosis H37Rv 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 cholesterol-degrading KshAMtb from Mycobacterium tuberculosis has the highest catalytic efficiency for CoA-thioesterified substrates, substrate specificity comparisons of KSH enzymes, overview Mycobacterium tuberculosis ?
-
 ?
1.14.15.30 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 ?
-
 ?
1.14.15.30 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 ?
-
 ?
1.14.15.30 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 ?
-
 ?
1.14.15.30 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 ?
-
 ?
1.14.15.30 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 ?
-
 ?
1.14.15.30 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
-
 ?
1.14.15.30 testosterone + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
 Rhodococcus rhodochrous (17beta)-9,17-dihydroxyandrost-4-en-3-one + oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.30 testosterone + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
 Rhodococcus rhodochrous ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.15.30 More KshAB is a two-component Rieske oxygenase comprising an oxygenase (KshA) and a reductase (KshB), structures comparisons of different KSH enzymes Rhodococcus rhodochrous
1.14.15.30 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
1.14.15.30 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

EC Number Synonyms Comment Organism
1.14.15.30 3-ketosteroid 9alpha-hydroxylase
-
 Rhodococcus rhodochrous
1.14.15.30 3-ketosteroid 9alpha-hydroxylase
-
 Mycobacterium tuberculosis
1.14.15.30 KshA
-
 Mycobacterium tuberculosis
1.14.15.30 KshA1
-
 Rhodococcus rhodochrous
1.14.15.30 KshA1 oxygenase component of 3-ketosteroid 9alpha-hydroxylase Rhodococcus rhodochrous
1.14.15.30 KshA5
-
 Rhodococcus rhodochrous
1.14.15.30 KshA5 oxygenase component of 3-ketosteroid 9alpha-hydroxylase Rhodococcus rhodochrous
1.14.15.30 KshAB
-
 Rhodococcus rhodochrous
1.14.15.30 KshAB
-
 Mycobacterium tuberculosis
1.14.15.30 KshAMt
-
 Mycobacterium tuberculosis
1.14.15.30 Rv3526
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.15.30 22
-
 assay at Rhodococcus rhodochrous
1.14.15.30 22
-
 assay at Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.15.30 0.5
-
 4-Estren-3,17-dione pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.5
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.5
-
 4-Estren-3,17-dione isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.5
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.6
-
 3-oxo-23,24-bisnorchol-4-en-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.6
-
 3-oxo-23,24-bisnorchol-4-en-22-oate isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.7
-
 androsta-1,4-diene-3,17-dione pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.7
-
 1,4-androstadiene-3,17-dione isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.8
-
 O2 pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.8
-
 testosterone pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.8
-
 4-Estren-3,17-dione pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.8
-
 O2 isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.8
-
 testosterone isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.8
-
 5alpha-androstan-3,17-dione isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.8
-
 4-Estren-3,17-dione isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 0.9
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.9
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 1.1
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 1.1
-
 3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 1.3
-
 androsta-1,4-diene-3,17-dione pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 1.3
-
 1,4-androstadiene-3,17-dione isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 1.4
-
 testosterone pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 1.4
-
 testosterone isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 1.7
-
 O2 pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 1.7
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 1.7
-
 O2 isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 1.7
-
 3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 2.6
-
 3-oxo-23,24-bisnorchol-4-en-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 2.6
-
 3-oxo-23,24-bisnorchol-4-en-22-oate isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.15.30 7
-
 assay at Rhodococcus rhodochrous
1.14.15.30 7
-
 assay at Mycobacterium tuberculosis

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.14.15.30 0.04
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.11
-
 3-oxo-23,24-bisnorchol-4-en-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 0.13
-
 testosterone pH 7.0, 22°C Rhodococcus rhodochrous

General Information

EC Number General Information Comment Organism
1.14.15.30 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
1.14.15.30 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
1.14.15.30 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
1.14.15.30 physiological function KshA is the oxygenase component of 3-ketosteroid 9alpha-hydroxylase, a Rieske oxygenase involved in the bacterial degradation of steroids Mycobacterium tuberculosis
1.14.15.30 physiological function KshA is the oxygenase component of 3-ketosteroid 9alpha-hydroxylase, a Rieske oxygenase involved in the bacterial degradation of steroids Rhodococcus rhodochrous
1.14.15.30 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 [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.15.30 1.2
-
 4-Estren-3,17-dione pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 1.2
-
 4-Estren-3,17-dione isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 3.7
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 3.7
-
 3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 13
-
 testosterone pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 13
-
 testosterone isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 17
-
 O2 pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 17
-
 O2 isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 27
-
 androsta-1,4-diene-3,17-dione pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 27
-
 1,4-androstadiene-3,17-dione isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 140
-
 5alpha-androstan-3,17-dione isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 500
-
 3-oxo-23,24-bisnorchol-4-en-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 500
-
 3-oxo-23,24-bisnorchol-4-en-22-oate isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 600
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 600
-
 3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 900
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 900
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 1000
-
 4-Estren-3,17-dione pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 1000
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 1000
-
 4-Estren-3,17-dione isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 1000
-
 3-oxo-23,24-bisnorchola-1,4-diene-22-oate isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 1100
-
 O2 pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 1100
-
 O2 isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 1200
-
 3-oxo-23,24-bisnorchol-4-en-22-oate pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 1200
-
 3-oxo-23,24-bisnorchol-4-en-22-oate isoform KshA1, at pH 7.0 and 22°C Rhodococcus rhodochrous
1.14.15.30 1500
-
 testosterone pH 7.0, 22°C Rhodococcus rhodochrous
1.14.15.30 1500
-
 testosterone isoform KshA5, at pH 7.0 and 22°C Rhodococcus rhodochrous