BRENDA - Enzyme Database

Substrate specificities and conformational flexibility of 3-ketosteroid 9alpha-hydroxylases

Penfield, J.S.; Worrall, L.J.; Strynadka, N.C.; Eltis, L.D.; J. Biol. Chem. 289, 25523-25536 (2014)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.14.15.30
expressed in Escherichia coli GJ1158 cells; expressed in Escherichia coli GJ1158 cells
Rhodococcus rhodochrous
Crystallization (Commentary)
EC Number
Crystallization
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; 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; 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; 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
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
Inhibitors
EC Number
Inhibitors
Commentary
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
Commentary
Organism
Structure
1.14.15.30
additional information
-
additional information
steady-state kinetic analysis
Mycobacterium tuberculosis
1.14.15.30
additional information
-
additional information
steady-state kinetic analysis; steady-state kinetic analysis
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; pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
0.0005
-
testosterone
isoform KshA5, at pH 7.0 and 22°C; 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; 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; 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; pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
0.0015
-
O2
isoform KshA1, at pH 7.0 and 22°C; 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
0.002
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
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; pH 7.0, 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
isoform KshA5, at pH 7.0 and 22°C; 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
0.05
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
0.11
-
testosterone
isoform KshA1, at pH 7.0 and 22°C; 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; 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
1.14.15.30
0.5
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
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
Rhodococcus rhodochrous DSM43269
-
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
Primary Accession No. (UniProt)
Commentary
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
; KshA1
-
1.14.15.30
Rhodococcus rhodochrous
F1CMY8
; KshA5
-
1.14.15.30
Rhodococcus rhodochrous DSM 43269
F1CMX0
KshA1
-
1.14.15.30
Rhodococcus rhodochrous DSM 43269
F1CMY8
KshA5
-
1.14.15.30
Rhodococcus rhodochrous DSM43269
F1CMX0
-
-
1.14.15.30
Rhodococcus rhodochrous DSM43269
F1CMY8
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.14.15.30
Ni-Sepharose column chromatography and Source 15Q column chromatography; Ni-Sepharose column chromatography and Source 15Q column chromatography
Rhodococcus rhodochrous
Reaction
EC Number
Reaction
Commentary
Organism
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
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; 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
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.15.30
1,4-androstadiene-3,17-dione + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
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
-
745308
Rhodococcus rhodochrous DSM43269
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
-
745308
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
-
745308
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
-
745308
Rhodococcus rhodochrous DSM43269
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
-
745308
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
-
745308
Rhodococcus rhodochrous
?
-
-
-
?
1.14.15.30
3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous DSM43269
?
-
-
-
?
1.14.15.30
3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
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
-
745308
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
745308
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
-
745308
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
-
745308
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
-
745308
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
-
745308
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
-
745308
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
-
745308
Rhodococcus rhodochrous
9alpha-hydroxy-4-estren-3,17-dione + oxidized ferredoxin [iron-sulfur] cluster + testosterone + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
?
-
-
?
1.14.15.30
5alpha-androstan-3,17-dione + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
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
-
745308
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
-
745308
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
-
745308
Rhodococcus rhodochrous DSM43269
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
-
745308
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
-
745308
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
745308
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
745308
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
745308
Rhodococcus rhodochrous
?
-
-
-
-
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
745308
Rhodococcus rhodochrous DSM43269
?
-
-
-
-
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
745308
Rhodococcus rhodochrous DSM43269
?
-
-
-
-
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
745308
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
745308
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
745308
Rhodococcus rhodochrous DSM 43269
?
-
-
-
-
1.14.15.30
testosterone + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
745308
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
-
745308
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
-
745308
Rhodococcus rhodochrous
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
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. 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
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; 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
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.14.15.30
22
-
assay at
Mycobacterium tuberculosis
1.14.15.30
22
-
assay at; assay at
Rhodococcus rhodochrous
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
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; 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; 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; 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
0.7
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
0.8
-
4-Estren-3,17-dione
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 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
-
O2
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
0.8
-
testosterone
isoform KshA5, at pH 7.0 and 22°C; 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; 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
1.1
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
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
1.3
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
1.4
-
testosterone
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 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
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; 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; pH 7.0, 22°C
Rhodococcus rhodochrous
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.14.15.30
7
-
assay at
Mycobacterium tuberculosis
1.14.15.30
7
-
assay at; assay at
Rhodococcus rhodochrous
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
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
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.15.30
expressed in Escherichia coli GJ1158 cells
Rhodococcus rhodochrous
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
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; 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
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; 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
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
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
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
Rhodococcus rhodochrous
1.14.15.30
4-androstene-3,17-dione
strong substrate inhibition
Rhodococcus rhodochrous
1.14.15.30
testosterone
substrate inhibition
Rhodococcus rhodochrous
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
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
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.15.30
additional information
-
additional information
steady-state kinetic analysis
Mycobacterium tuberculosis
1.14.15.30
additional information
-
additional information
steady-state kinetic analysis
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; pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
0.0005
-
testosterone
isoform KshA5, at pH 7.0 and 22°C; 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; 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; 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; pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
0.0015
-
O2
isoform KshA1, at pH 7.0 and 22°C; 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
0.002
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
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; pH 7.0, 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
isoform KshA5, at pH 7.0 and 22°C; 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
0.05
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
0.11
-
testosterone
isoform KshA1, at pH 7.0 and 22°C; 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; 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
1.14.15.30
0.5
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
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
Rhodococcus rhodochrous DSM43269
-
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
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.15.30
Ni-Sepharose column chromatography and Source 15Q column chromatography
Rhodococcus rhodochrous
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.15.30
1,4-androstadiene-3,17-dione + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
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
-
745308
Rhodococcus rhodochrous DSM43269
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
-
745308
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
-
745308
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
-
745308
Rhodococcus rhodochrous DSM43269
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
-
745308
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
-
745308
Rhodococcus rhodochrous
?
-
-
-
?
1.14.15.30
3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous DSM43269
?
-
-
-
?
1.14.15.30
3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
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
-
745308
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
745308
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
-
745308
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
-
745308
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
-
745308
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
-
745308
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
-
745308
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
-
745308
Rhodococcus rhodochrous
9alpha-hydroxy-4-estren-3,17-dione + oxidized ferredoxin [iron-sulfur] cluster + testosterone + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
?
-
-
?
1.14.15.30
5alpha-androstan-3,17-dione + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
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
-
745308
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
-
745308
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
-
745308
Rhodococcus rhodochrous DSM43269
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
-
745308
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
-
745308
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
745308
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
745308
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
745308
Rhodococcus rhodochrous
?
-
-
-
-
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
745308
Rhodococcus rhodochrous DSM43269
?
-
-
-
-
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
745308
Rhodococcus rhodochrous DSM43269
?
-
-
-
-
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
745308
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
745308
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
745308
Rhodococcus rhodochrous DSM 43269
?
-
-
-
-
1.14.15.30
testosterone + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
745308
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
-
745308
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
-
745308
Rhodococcus rhodochrous
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
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. 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
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
Rhodococcus rhodochrous
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.14.15.30
22
-
assay at
Mycobacterium tuberculosis
1.14.15.30
22
-
assay at
Rhodococcus rhodochrous
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
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; 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; 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; 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
0.7
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
0.8
-
4-Estren-3,17-dione
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 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
-
O2
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
0.8
-
testosterone
isoform KshA5, at pH 7.0 and 22°C; 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; 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
1.1
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
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
1.3
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
1.4
-
testosterone
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 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
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; 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; pH 7.0, 22°C
Rhodococcus rhodochrous
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.14.15.30
7
-
assay at
Mycobacterium tuberculosis
1.14.15.30
7
-
assay at
Rhodococcus rhodochrous
General Information
EC Number
General Information
Commentary
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
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. 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; 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; 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
General Information (protein specific)
EC Number
General Information
Commentary
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
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. 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. 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. KshA1 has a role in bile acid catabolism
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
Rhodococcus rhodochrous
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.14.15.30
1.2
-
4-Estren-3,17-dione
isoform KshA1, at pH 7.0 and 22°C; 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
3.7
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
13
-
testosterone
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
17
-
O2
isoform KshA5, at pH 7.0 and 22°C; 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
27
-
androsta-1,4-diene-3,17-dione
pH 7.0, 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
isoform KshA5, at pH 7.0 and 22°C; 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
600
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
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; pH 7.0, 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; 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; pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
1100
-
O2
isoform KshA1, at pH 7.0 and 22°C; 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; pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
1500
-
testosterone
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.14.15.30
1.2
-
4-Estren-3,17-dione
isoform KshA1, at pH 7.0 and 22°C; 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
3.7
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
13
-
testosterone
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
17
-
O2
isoform KshA5, at pH 7.0 and 22°C; 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
27
-
androsta-1,4-diene-3,17-dione
pH 7.0, 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
isoform KshA5, at pH 7.0 and 22°C; 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
600
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
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; pH 7.0, 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; 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; pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
1100
-
O2
isoform KshA1, at pH 7.0 and 22°C; 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; pH 7.0, 22°C
Rhodococcus rhodochrous
1.14.15.30
1500
-
testosterone
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous