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Literature summary extracted from

  • Capyk, J.; Casabon, I.; Gruninger, R.; Strynadka, N.; Eltis, L.
    Activity of 3-ketosteroid 9alpha-hydroxylase (KshAB) indicates cholesterol side chain and ring degradation occur simultaneously in Mycobacterium tuberculosis (2011), J. Biol. Chem., 286, 40717-40724.
    View publication on PubMedView publication on EuropePMC

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.15.30 additional information
-
 additional information steady-state kinetic parameters for KshAB, overview Mycobacterium tuberculosis
1.14.15.30 0.003
-
 3-oxo-23,24-bisnorchol-4-en-22-oic acid in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 0.003
-
 3-oxo-23,24-bisnorcholesta-4-ene-22-oate pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 0.0068
-
 3-oxo-23,24-bisnorchol-4-en-22-oic acid-CoA in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 0.0068
-
 3-oxo-23,24-bisnorcholesta-4-ene-22-oyl-CoA pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 0.017
-
 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid-CoA in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 0.017
-
 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate-CoA pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 0.024
-
 4-androstene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 0.024
-
 4-androstene-3,17-dione pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 0.07
-
 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 0.07
-
 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 0.09
-
 O2 apparent Km value, in the presence of 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid-CoA, in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 0.1 1 androsta-1,4-diene-3,17-dione pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 0.11
-
 androsta-1,4-diene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.15.30 3-oxo-23,24-bisnorcholest-4-en-22-oyl-CoA + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 Mycobacterium tuberculosis
-
 9alpha-hydroxy-3-oxo-23,24-bisnorcholest-4-en-22-oyl-CoA + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.30 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oyl-CoA + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 Mycobacterium tuberculosis
-
 9alpha-hydroxy-3-oxo-23,24-bisnorcholesta-1,4-dien-22-oyl-CoA + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.15.30 Mycobacterium tuberculosis
-
-
-
1.14.15.30 Mycobacterium tuberculosis P71875 gene kshA; the enzyme harbors a Rieske oxygenase, encoded by kshA
-
1.14.15.30 Mycobacterium tuberculosis P9WJ93 gene kshB; the enzyme harbors a reductase, encoded by kshB
-
1.14.15.30 Mycobacterium tuberculosis H37Rv P71875 gene kshA; the enzyme harbors a Rieske oxygenase, encoded by kshA
-
1.14.15.30 Mycobacterium tuberculosis H37Rv P9WJ93 gene kshB; the enzyme harbors a reductase, encoded by kshB
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.15.30 3-oxo-23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
 Mycobacterium tuberculosis 9alpha-hydroxy-3-oxo-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.30 3-oxo-23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
 Mycobacterium tuberculosis H37Rv 9alpha-hydroxy-3-oxo-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.30 3-oxo-23,24-bisnorcholest-4-en-22-oyl-CoA + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
 Mycobacterium tuberculosis 9alpha-hydroxy-3-oxo-23,24-bisnorcholest-4-en-22-oyl-CoA + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.30 3-oxo-23,24-bisnorcholest-4-en-22-oyl-CoA + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
 Mycobacterium tuberculosis H37Rv 9alpha-hydroxy-3-oxo-23,24-bisnorcholest-4-en-22-oyl-CoA + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.30 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
 Mycobacterium tuberculosis 9alpha-hydroxy-3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.30 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
 Mycobacterium tuberculosis H37Rv 9alpha-hydroxy-3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.30 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate-CoA + reduced ferredoxin [iron-sulfur] cluster + H+ + O2
-
 Mycobacterium tuberculosis ?
-
 r
1.14.15.30 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oyl-CoA + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
 Mycobacterium tuberculosis 9alpha-hydroxy-3-oxo-23,24-bisnorcholesta-1,4-dien-22-oyl-CoA + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.30 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oyl-CoA + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
 Mycobacterium tuberculosis H37Rv 9alpha-hydroxy-3-oxo-23,24-bisnorcholesta-1,4-dien-22-oyl-CoA + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.30 4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
 Mycobacterium tuberculosis 9alpha-hydroxy-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.30 4-androstene-3,17-dione + O2 + NADH + H+
-
 Mycobacterium tuberculosis ? + H2O + NAD+
-
 ?
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 additional information substrate specificity, overview. KshAB also transforms CoA thioester steroids Mycobacterium tuberculosis ?
-
 ?
1.14.15.30 additional information substrate specificity, overview. KshAB also transforms CoA thioester steroids Mycobacterium tuberculosis H37Rv ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.15.30 More the enzyme comprises a Rieske oxygenase, encoded by kshA, and a reductase, encoded by kshB Mycobacterium tuberculosis

Synonyms

EC Number Synonyms Comment Organism
1.14.15.30 3-ketosteroid 9alpha-hydroxylase
-
 Mycobacterium tuberculosis
1.14.15.30 KshAB
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

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

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.15.30 0.07
-
 4-androstene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 0.07
-
 4-androstene-3,17-dione pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 0.08
-
 3-oxo-23,24-bisnorchol-4-en-22-oic acid in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 0.08
-
 3-oxo-23,24-bisnorcholesta-4-ene-22-oate pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 0.25
-
 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 0.25
-
 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 0.61
-
 3-oxo-23,24-bisnorchol-4-en-22-oic acid-CoA in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 0.61
-
 3-oxo-23,24-bisnorcholesta-4-ene-22-oyl-CoA pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 0.8
-
 androsta-1,4-diene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 0.8
-
 androsta-1,4-diene-3,17-dione pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 2.5
-
 O2 apparent Km value, in the presence of 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid-CoA, in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 2.7
-
 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid-CoA in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 2.7
-
 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate-CoA pH 7.0, 22°C, KshAB Mycobacterium tuberculosis

pH Optimum

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

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.15.30 Fe-S center Rieske-type Fe-S cluster, Rieske monooxygenase Mycobacterium tuberculosis

General Information

EC Number General Information Comment Organism
1.14.15.30 additional information enzyme substrate binding pocket analysis using molecular modeling, overview. Modeling of substrates into KshA crystal structure using KshA structure PDB 2ZYL Mycobacterium tuberculosis
1.14.15.30 physiological function the enzyme plays a role of cholesterol catabolism, overview Mycobacterium tuberculosis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.15.30 2.917
-
 4-androstene-3,17-dione pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 3
-
 4-androstene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 3.5
-
 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 3.57
-
 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 7.27
-
 androsta-1,4-diene-3,17-dione pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 7.6
-
 androsta-1,4-diene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 26.67
-
 3-oxo-23,24-bisnorcholesta-4-ene-22-oate pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 29
-
 O2 apparent Km value, in the presence of 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid-CoA, in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 30
-
 3-oxo-23,24-bisnorchol-4-en-22-oic acid in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 89.71
-
 3-oxo-23,24-bisnorcholesta-4-ene-22-oyl-CoA pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 90
-
 3-oxo-23,24-bisnorchol-4-en-22-oic acid-CoA in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis
1.14.15.30 158.8
-
 3-oxo-23,24-bisnorcholesta-1,4-dien-22-oate-CoA pH 7.0, 22°C, KshAB Mycobacterium tuberculosis
1.14.15.30 160
-
 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid-CoA in 0.1 M potassium phosphate, pH 7.0, at 22°C Mycobacterium tuberculosis