BRENDA - Enzyme Database
show all sequences of 1.14.15.30

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)
Commentary
Organism
expressed in Escherichia coli GJ1158 cells; expressed in Escherichia coli GJ1158 cells
Rhodococcus rhodochrous
Crystallization (Commentary)
Crystallization
Organism
in complex with 1,4-androstadiene-3,17-dione or 3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA, hanging drop vapor diffusion method, using 500 mM NaH2PO4, 125 mM K2HPO4, 4% (w/v) PEG-1000 or 2% (w/v) PEG-3000, 20 mM Tris, 100 mM phosphate-citrate, pH 4.2; 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
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
Inhibitors
Commentary
Organism
Structure
3-oxo-23,24-bisnorchol-4-en-22-oate
substrate inhibition
Rhodococcus rhodochrous
3-oxo-23,24-bisnorchola-1,4-diene-22-oate
strong substrate inhibition; strong substrate inhibition
Rhodococcus rhodochrous
4-androstene-3,17-dione
strong substrate inhibition; strong substrate inhibition
Rhodococcus rhodochrous
testosterone
substrate inhibition
Rhodococcus rhodochrous
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
steady-state kinetic analysis
Mycobacterium tuberculosis
additional information
-
additional information
steady-state kinetic analysis; steady-state kinetic analysis
Rhodococcus rhodochrous
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
0.0005
-
testosterone
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.0008
-
4-Estren-3,17-dione
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.001
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oate
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.0012
-
3-oxo-23,24-bisnorchol-4-en-22-oate
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.0015
-
O2
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.002
-
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
0.002
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
0.0022
-
3-oxo-23,24-bisnorchol-4-en-22-oate
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.0061
-
5alpha-Androstan-3,17-dione
isoform KshA5, at pH 7.0 and 22°C
Rhodococcus rhodochrous
0.049
-
O2
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.05
-
1,4-androstadiene-3,17-dione
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
0.05
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
0.11
-
testosterone
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.4
-
4-Estren-3,17-dione
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.5
-
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
0.5
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Rhodococcus rhodochrous
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Mycobacterium tuberculosis
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Rhodococcus rhodochrous DSM43269
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Mycobacterium tuberculosis H37Rv
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Rhodococcus rhodochrous DSM 43269
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Mycobacterium tuberculosis
P71875
KshA
-
Mycobacterium tuberculosis H37Rv
P71875
KshA
-
Rhodococcus rhodochrous
F1CMX0
; KshA1
-
Rhodococcus rhodochrous
F1CMY8
; KshA5
-
Rhodococcus rhodochrous DSM 43269
F1CMX0
KshA1
-
Rhodococcus rhodochrous DSM 43269
F1CMY8
KshA5
-
Rhodococcus rhodochrous DSM43269
F1CMX0
-
-
Rhodococcus rhodochrous DSM43269
F1CMY8
-
-
Purification (Commentary)
Commentary
Organism
Ni-Sepharose column chromatography and Source 15Q column chromatography; Ni-Sepharose column chromatography and Source 15Q column chromatography
Rhodococcus rhodochrous
Reaction
Reaction
Commentary
Organism
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
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)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
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
-
-
-
?
1,4-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
-
-
-
?
1,4-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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous
?
-
-
-
?
3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous DSM43269
?
-
-
-
?
3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous DSM 43269
?
-
-
-
?
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous
?
-
-
-
?
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
-
-
-
?
3-oxo-23,24-bisnorchola-1,4-diene-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous
?
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
?
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
?
-
-
-
-
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
?
-
-
-
-
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
?
-
-
-
-
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
?
-
-
-
-
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
?
-
-
-
-
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
?
-
-
-
-
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
?
-
-
-
-
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
?
-
-
-
-
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
-
-
-
?
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
-
-
-
?
testosterone + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous
?
-
-
-
?
Subunits
Subunits
Commentary
Organism
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
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]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
22
-
assay at
Mycobacterium tuberculosis
22
-
assay at; assay at
Rhodococcus rhodochrous
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
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
0.5
-
4-Estren-3,17-dione
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.6
-
3-oxo-23,24-bisnorchol-4-en-22-oate
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.7
-
1,4-androstadiene-3,17-dione
isoform KshA5, at pH 7.0 and 22°C
Rhodococcus rhodochrous
0.7
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
0.8
-
4-Estren-3,17-dione
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.8
-
5alpha-Androstan-3,17-dione
isoform KshA5, at pH 7.0 and 22°C
Rhodococcus rhodochrous
0.8
-
O2
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.8
-
testosterone
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.9
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oate
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1.1
-
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
1.1
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
1.3
-
1,4-androstadiene-3,17-dione
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
1.3
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
1.4
-
testosterone
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1.7
-
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
1.7
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
1.7
-
O2
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
2.6
-
3-oxo-23,24-bisnorchol-4-en-22-oate
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Mycobacterium tuberculosis
7
-
assay at; assay at
Rhodococcus rhodochrous
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.04
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oate
pH 7.0, 22°C
Rhodococcus rhodochrous
0.11
-
3-oxo-23,24-bisnorchol-4-en-22-oate
pH 7.0, 22°C
Rhodococcus rhodochrous
0.13
-
testosterone
pH 7.0, 22°C
Rhodococcus rhodochrous
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli GJ1158 cells
Rhodococcus rhodochrous
Crystallization (Commentary) (protein specific)
Crystallization
Organism
in complex with 1,4-androstadiene-3,17-dione or 3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA, hanging drop vapor diffusion method, using 500 mM NaH2PO4, 125 mM K2HPO4, 4% (w/v) PEG-1000 or 2% (w/v) PEG-3000, 20 mM Tris, 100 mM phosphate-citrate, pH 4.2; 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
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
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)
Inhibitors
Commentary
Organism
Structure
3-oxo-23,24-bisnorchol-4-en-22-oate
substrate inhibition
Rhodococcus rhodochrous
3-oxo-23,24-bisnorchola-1,4-diene-22-oate
strong substrate inhibition
Rhodococcus rhodochrous
4-androstene-3,17-dione
strong substrate inhibition
Rhodococcus rhodochrous
testosterone
substrate inhibition
Rhodococcus rhodochrous
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.04
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oate
pH 7.0, 22°C
Rhodococcus rhodochrous
0.11
-
3-oxo-23,24-bisnorchol-4-en-22-oate
pH 7.0, 22°C
Rhodococcus rhodochrous
0.13
-
testosterone
pH 7.0, 22°C
Rhodococcus rhodochrous
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
steady-state kinetic analysis
Mycobacterium tuberculosis
additional information
-
additional information
steady-state kinetic analysis
Rhodococcus rhodochrous
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
0.0005
-
testosterone
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.0008
-
4-Estren-3,17-dione
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.001
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oate
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.0012
-
3-oxo-23,24-bisnorchol-4-en-22-oate
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.0015
-
O2
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.002
-
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
0.002
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
0.0022
-
3-oxo-23,24-bisnorchol-4-en-22-oate
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.0061
-
5alpha-Androstan-3,17-dione
isoform KshA5, at pH 7.0 and 22°C
Rhodococcus rhodochrous
0.049
-
O2
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.05
-
1,4-androstadiene-3,17-dione
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
0.05
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
0.11
-
testosterone
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.4
-
4-Estren-3,17-dione
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.5
-
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
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)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Rhodococcus rhodochrous
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Mycobacterium tuberculosis
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Rhodococcus rhodochrous DSM43269
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Mycobacterium tuberculosis H37Rv
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
Rhodococcus rhodochrous DSM 43269
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
Ni-Sepharose column chromatography and Source 15Q column chromatography
Rhodococcus rhodochrous
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
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
-
-
-
?
1,4-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
-
-
-
?
1,4-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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous
?
-
-
-
?
3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous DSM43269
?
-
-
-
?
3-oxo-23,24-bisnorchol-4-en-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous DSM 43269
?
-
-
-
?
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous
?
-
-
-
?
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
-
-
-
?
3-oxo-23,24-bisnorchola-1,4-diene-22-oate + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous
?
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
?
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
?
-
-
-
-
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
?
-
-
-
-
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
?
-
-
-
-
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
?
-
-
-
-
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
?
-
-
-
-
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
?
-
-
-
-
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
?
-
-
-
-
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
?
-
-
-
-
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
-
-
-
?
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
-
-
-
?
testosterone + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
745308
Rhodococcus rhodochrous
?
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
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
More
KshAB is a two-component Rieske oxygenase comprising an oxygenase (KshA) and a reductase (KshB), structures comparisons of different KSH enzymes. The substrate binding residues within the active site pocket are positioned nearly identically in enzyme KshA1 from Rhodococcus rhodochrous and KshA from Mycobacterium tuberculosis
Rhodococcus rhodochrous
More
KshAB is a two-component Rieske oxygenase comprising an oxygenase (KshA) and a reductase (KshB), structures comparisons of different KSH enzymes
Rhodococcus rhodochrous
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
22
-
assay at
Mycobacterium tuberculosis
22
-
assay at
Rhodococcus rhodochrous
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
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
0.5
-
4-Estren-3,17-dione
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.6
-
3-oxo-23,24-bisnorchol-4-en-22-oate
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.7
-
1,4-androstadiene-3,17-dione
isoform KshA5, at pH 7.0 and 22°C
Rhodococcus rhodochrous
0.7
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
0.8
-
4-Estren-3,17-dione
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.8
-
5alpha-Androstan-3,17-dione
isoform KshA5, at pH 7.0 and 22°C
Rhodococcus rhodochrous
0.8
-
O2
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.8
-
testosterone
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
0.9
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oate
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1.1
-
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
1.1
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
1.3
-
1,4-androstadiene-3,17-dione
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
1.3
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
1.4
-
testosterone
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1.7
-
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
1.7
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
1.7
-
O2
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
2.6
-
3-oxo-23,24-bisnorchol-4-en-22-oate
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Mycobacterium tuberculosis
7
-
assay at
Rhodococcus rhodochrous
General Information
General Information
Commentary
Organism
additional information
the enzyme has a steroid-binding pocket with the substrate's C17 oriented toward the active site opening. Comparisons suggest that Tyr245 and Phe297 are determinants of KshA1 specificity. The enzyme has a flexible 16-residue mouth loop, which in some structures completely occlude the substrate-binding pocket from the bulk solvent
Mycobacterium tuberculosis
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
physiological function
KshA is the oxygenase component of 3-ketosteroid 9alpha-hydroxylase, a Rieske oxygenase involved in the bacterial degradation of steroids
Mycobacterium tuberculosis
physiological function
KshA is the oxygenase component of 3-ketosteroid 9alpha-hydroxylase, a Rieske oxygenase involved in the bacterial degradation of steroids; 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)
General Information
Commentary
Organism
additional information
the enzyme has a steroid-binding pocket with the substrate's C17 oriented toward the active site opening. Comparisons suggest that Tyr245 and Phe297 are determinants of KshA1 specificity. The enzyme has a flexible 16-residue mouth loop, which in some structures completely occlude the substrate-binding pocket from the bulk solvent
Mycobacterium tuberculosis
additional information
the enzyme has a steroid-binding pocket with the substrate's C17 oriented toward the active site opening. Comparisons suggest that Tyr245 and Phe297 are determinants of KshA1 specificity. The enzyme has a flexible 16-residue mouth loop, which in some structures completely occlude the substrate-binding pocket from the bulk solvent
Rhodococcus rhodochrous
additional information
the enzyme has a steroid-binding pocket with the substrate's C17 oriented toward the active site opening. The enzyme has a flexible 16-residue mouth loop, which in some structures completely occlude the substrate-binding pocket from the bulk solvent. The catalytic iron andalpha-helices harboring its ligands are displaced up to 4.4 A in theKshA5substrate complexes as compared with substrate-free KshA, suggesting that Rieske oxygenases may have a dynamic nature similar to cytochrome P450
Rhodococcus rhodochrous
physiological function
KshA is the oxygenase component of 3-ketosteroid 9alpha-hydroxylase, a Rieske oxygenase involved in the bacterial degradation of steroids
Mycobacterium tuberculosis
physiological function
KshA is the oxygenase component of 3-ketosteroid 9alpha-hydroxylase, a Rieske oxygenase involved in the bacterial degradation of steroids. KshA1 has a role in bile acid catabolism
Rhodococcus rhodochrous
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]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.2
-
4-Estren-3,17-dione
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
3.7
-
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
3.7
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
13
-
testosterone
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
17
-
O2
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
27
-
1,4-androstadiene-3,17-dione
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
27
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
140
-
5alpha-Androstan-3,17-dione
isoform KshA5, at pH 7.0 and 22°C
Rhodococcus rhodochrous
500
-
3-oxo-23,24-bisnorchol-4-en-22-oate
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
600
-
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
600
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
900
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oate
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
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
1000
-
4-Estren-3,17-dione
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1100
-
O2
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1200
-
3-oxo-23,24-bisnorchol-4-en-22-oate
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1500
-
testosterone
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.2
-
4-Estren-3,17-dione
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
3.7
-
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
3.7
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
13
-
testosterone
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
17
-
O2
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
27
-
1,4-androstadiene-3,17-dione
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
27
-
androsta-1,4-diene-3,17-dione
pH 7.0, 22°C
Rhodococcus rhodochrous
140
-
5alpha-Androstan-3,17-dione
isoform KshA5, at pH 7.0 and 22°C
Rhodococcus rhodochrous
500
-
3-oxo-23,24-bisnorchol-4-en-22-oate
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
600
-
3-oxo-23,24-bisnorchola-1,4-dien-22-oate-CoA
isoform KshA1, at pH 7.0 and 22°C
Rhodococcus rhodochrous
600
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oyl-CoA
pH 7.0, 22°C
Rhodococcus rhodochrous
900
-
3-oxo-23,24-bisnorchola-1,4-diene-22-oate
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
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
1000
-
4-Estren-3,17-dione
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1100
-
O2
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1200
-
3-oxo-23,24-bisnorchol-4-en-22-oate
isoform KshA1, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
1500
-
testosterone
isoform KshA5, at pH 7.0 and 22°C; pH 7.0, 22°C
Rhodococcus rhodochrous
Other publictions for EC 1.14.15.30
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
745648
Guevara
Functional characterization o ...
Rhodococcus ruber, Rhodococcus ruber Chol-4
J. Steroid Biochem. Mol. Biol.
172
176-187
2017
-
-
1
-
1
-
-
-
-
-
-
4
-
6
-
-
-
-
-
-
-
-
22
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
22
1
-
-
-
-
-
-
-
-
-
4
4
-
-
-
746503
Zhang
Efficient 9alpha-hydroxy-4-an ...
Mycolicibacterium neoaurum, Mycolicibacterium neoaurum JC-12
SpringerPlus
5
1207
2016
-
-
1
-
1
-
5
-
-
-
-
8
-
2
-
-
1
-
-
-
1
1
10
2
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
1
-
-
5
-
-
-
-
-
8
-
-
-
1
-
-
1
1
10
2
1
-
-
-
1
-
-
-
-
1
1
-
-
-
743221
Yao
Characterization and engineer ...
Mycolicibacterium neoaurum, Mycolicibacterium neoaurum ATCC 25795, Mycolicibacterium neoaurum NwIB-01
Metab. Eng.
24
181-191
2014
-
-
1
-
1
-
-
-
-
1
-
3
-
4
-
-
-
-
-
-
-
-
3
-
1
-
-
-
1
-
-
1
-
-
-
-
-
1
1
-
1
-
-
-
-
-
-
1
-
3
-
-
-
-
-
-
-
-
3
-
1
-
-
-
1
-
-
-
1
-
-
1
-
-
744058
Petrusma
3-Ketosteroid 9alpha-hydroxyl ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv, Mycolicibacterium smegmatis, Mycolicibacterium smegmatis mc2 155, Rhodococcus erythropolis, Rhodococcus erythropolis SQ1, Rhodococcus jostii, Rhodococcus rhodochrous, Rhodococcus rhodochrous DSM 43269
Antonie van Leeuwenhoek
106
157-172
2014
-
2
2
-
1
-
-
-
-
5
-
22
-
15
-
-
-
5
-
-
-
-
28
5
-
-
-
-
-
-
-
10
-
-
-
-
2
4
14
-
1
-
-
-
-
-
-
7
-
22
-
-
-
-
-
-
-
-
28
7
-
-
-
-
-
-
-
-
-
20
28
-
-
-
745308
Penfield
Substrate specificities and c ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv, Rhodococcus rhodochrous, Rhodococcus rhodochrous DSM43269, Rhodococcus rhodochrous DSM 43269
J. Biol. Chem.
289
25523-25536
2014
-
-
1
2
-
-
4
19
-
-
-
8
-
18
-
-
1
2
-
-
-
-
54
2
2
-
-
19
2
-
-
-
3
-
-
-
-
2
-
3
-
-
-
6
3
20
-
-
-
8
-
-
-
2
-
-
-
-
54
3
3
-
-
19
3
-
-
-
-
4
6
-
17
17
745698
Yeh
Deletion of the gene encoding ...
Rhodococcus hoagii 103S, Rhodococcus hoagii
Microb. Cell Fact.
13
130
2014
-
-
1
-
1
-
-
-
-
-
-
4
-
2
-
-
-
-
-
-
-
-
4
-
1
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
4
-
1
-
-
-
-
-
-
-
-
1
1
-
-
-
717737
Petrusma
Structural features in the Ksh ...
Rhodococcus rhodochrous, Rhodococcus rhodochrous DSM 43269
J. Bacteriol.
194
115-121
2012
-
-
1
-
8
-
-
2
-
1
-
3
-
13
-
-
1
-
-
-
-
-
74
-
-
-
-
-
1
-
-
2
-
-
-
-
-
3
4
-
16
-
-
-
-
4
-
2
-
3
-
-
-
3
-
-
-
-
74
-
-
-
-
-
2
-
-
-
-
1
2
-
-
-
717735
Petrusma
Multiplicity of 3-ketosteroid- ...
Rhodococcus rhodochrous, Rhodococcus rhodochrous DSM 43269
J. Bacteriol.
193
3931-3940
2011
-
-
1
-
1
-
-
-
-
-
-
4
-
19
-
-
1
-
-
1
-
-
75
-
-
-
-
-
1
-
-
1
-
-
-
-
-
6
5
-
5
-
-
-
-
-
-
-
-
4
-
-
-
6
-
3
-
-
75
-
-
-
-
-
3
-
-
-
1
3
11
5
-
-
717854
Capyk
Activity of 3-ketosteroid 9alp ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Biol. Chem.
286
40717-40724
2011
-
-
-
-
-
-
-
13
-
-
-
2
-
9
-
-
-
-
-
-
-
-
30
1
1
-
-
11
1
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
21
-
-
-
2
-
-
-
-
-
-
-
-
30
2
2
-
-
19
2
-
-
-
-
2
3
-
13
19
713784
Wei
A new steroid-transforming str ...
Mycolicibacterium neoaurum, Mycolicibacterium neoaurum NwIB-01
Appl. Biochem. Biotechnol.
162
1446-1456
2010
-
-
-
-
-
-
-
-
-
-
1
-
-
3
-
-
-
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
2
1
-
-
-
-
-
-
-
1
-
1
1
-
-
-
716250
Hu
3-Ketosteroid 9alpha-hydroxyla ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Mol. Microbiol.
75
107-121
2010
-
-
-
-
-
-
-
-
-
-
-
-
-
160
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
717500
Fan
Cloning, heterologous expressi ...
Mycobacterium sp., Mycobacterium sp. NwIB-01
Chin. J. Biotechnol.
25
2014-2021
2009
-
-
1
-
-
-
-
-
-
-
1
-
-
8
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
717784
Capyk
Characterization of 3-ketoster ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Biol. Chem.
284
9937-9946
2009
-
-
1
1
-
-
-
3
-
-
-
-
-
6
-
-
1
-
-
-
-
-
4
-
-
-
-
3
-
-
-
1
-
-
-
-
-
1
1
1
-
-
-
-
-
3
-
-
-
-
-
-
-
1
-
-
-
-
4
-
-
-
-
3
-
-
-
-
-
-
-
-
3
3
717119
Van Der Geize
Characterization of a second R ...
Rhodococcus erythropolis, Rhodococcus erythropolis SQ1
Appl. Environ. Microbiol.
74
7197-7203
2008
-
-
-
-
-
-
-
-
-
-
-
-
-
9
-
-
-
-
-
-
-
-
8
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
8
-
-
-
-
-
-
-
-
-
1
2
3
1
-
-
717118
Andor
Generation of useful insertion ...
Mycolicibacterium smegmatis, Mycolicibacterium smegmatis mc(2)155 / ATCC 700084
Appl. Environ. Microbiol.
72
6554-6559
2006
-
-
1
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
718125
Van Der Geize
Molecular and functional chara ...
Rhodococcus erythropolis, Rhodococcus erythropolis SQ1
Mol. Microbiol.
45
1007-1018
2002
-
-
-
-
1
-
-
-
-
1
-
-
-
4
-
-
-
-
-
-
-
-
8
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
8
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-