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Literature summary for 1.14.15.30 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
Search for more literature for 1.14.15.30

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.024
-
 4-androstene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis
0.11
-
 1,4-androstadiene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis
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

Organism UniProt Comment Textmining
Mycobacterium tuberculosis P71875
-
-
Mycobacterium tuberculosis H37Rv P71875
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.07
-
 4-androstene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis
0.8
-
 1,4-androstadiene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis
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

Cofactor Comment Organism Structure
[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]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
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
3
-
 4-androstene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis
7.6
-
 1,4-androstadiene-3,17-dione in 0.1 M potassium phosphate, pH 7.0, at 25°C Mycobacterium tuberculosis