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Literature summary extracted from

  • Capyk, J.; DAngelo, I.; Strynadka, N.; Eltis, L.
    Characterization of 3-ketosteroid 9alpha-hydroxylase, a Rieske oxygenase in the cholesterol degradation pathway of Mycobacterium tuberculosis (2009), J. Biol. Chem., 284, 9937-9946.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.15.30 expressed in Escherichia coli BL21(DE3) cells Mycobacterium tuberculosis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.14.15.30 component KshA, sitting drop vapor diffusion method, using 1.1 M sodium malonate, 0.1 M HEPES, pH 7.0, and 0.5% (v/v) Jeffamine M600 Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.15.30 0.024
-
 4-androstene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis
1.14.15.30 0.11
-
 1,4-androstadiene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis
1.14.15.30 1.2
-
 O2 in the presence of 1,4-androstadiene-3,17-dione, Km above 1.2 mM, in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
1.14.15.30 Mycobacterium tuberculosis P71875
-
-
1.14.15.30 Mycobacterium tuberculosis H37Rv P71875
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.15.30 Ni2+-NTA agarose column chromatography and Source15Q column chromatography Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.15.30 4-androstene-3,17-dione + O2 + NADH + H+
-
 Mycobacterium tuberculosis 9alpha-hydroxy-4-androstene-3,17-dione + H2O + NAD+
-
 ?
1.14.15.30 4-androstene-3,17-dione + O2 + NADH + H+
-
 Mycobacterium tuberculosis H37Rv 9alpha-hydroxy-4-androstene-3,17-dione + H2O + NAD+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.14.15.30 3-ketosteroid 9alpha-hydroxylase
-
 Mycobacterium tuberculosis
1.14.15.30 KshAB
-
 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 25°C Mycobacterium tuberculosis
1.14.15.30 0.8
-
 1,4-androstadiene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis
1.14.15.30 7
-
 O2 in the presence of 1,4-androstadiene-3,17-dione, kcat above 7 1/sec, in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.15.30 [2Fe-2S]-center 8.2 mol of iron and 2.4 mol of sulfur per mol of KshA protomer 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 2.45
-
 O2 in the presence of 1,4-androstadiene-3,17-dione, in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis
1.14.15.30 3
-
 4-androstene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis
1.14.15.30 7.6
-
 1,4-androstadiene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis