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Literature summary extracted from

  • Ouellet, H.; Guan, S.; Johnston, J.; Chow, E.; Kells, P.; Burlingame, A.; Cox, J.; Podust, L.; De Montellano, P.
    Mycobacterium tuberculosis CYP125A1, a steroid C27 monooxygenase that detoxifies intracellularly generated cholest-4-en-3-one (2010), Mol. Microbiol., 77, 730-742.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.15.29 expressed in Escherichia coli DH5alpha cells Mycobacterium tuberculosis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.14.15.29 enzyme bound to cholest-4-en-3-one, hanging drop vapor diffusion method, using 0.2 M ammonium sulfate, 0.1 M bis-Tris, pH 5.5, and 25% (w/v) PEG 3350 Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.15.29 27-hydroxycholest-4-en-3-one + 4 reduced ferredoxin [iron-sulfur] cluster + 4 H+ + O2 Mycobacterium tuberculosis
-
 cholest-4-en-3-one-27-oic acid + 4 oxidized ferredoxin [iron-sulfur] cluster + 3 H2O
-
 ?
1.14.15.29 27-hydroxycholest-4-en-3-one + 4 reduced ferredoxin [iron-sulfur] cluster + 4 H+ + O2 Mycobacterium tuberculosis CDC 1551
-
 cholest-4-en-3-one-27-oic acid + 4 oxidized ferredoxin [iron-sulfur] cluster + 3 H2O
-
 ?
1.14.15.29 cholest-4-en-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 Mycobacterium tuberculosis physiological substrate, the enzyme hydroxylates cholest-4-en-3-one at C-27 27-hydroxycholest-4-en-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.29 cholest-4-en-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 Mycobacterium tuberculosis CDC 1551 physiological substrate, the enzyme hydroxylates cholest-4-en-3-one at C-27 27-hydroxycholest-4-en-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.15.29 Mycobacterium tuberculosis P9WPP0
-
-
1.14.15.29 Mycobacterium tuberculosis CDC 1551 P9WPP0
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.15.29
-
 Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.15.29 27-hydroxycholest-4-en-3-one + 4 reduced ferredoxin [iron-sulfur] cluster + 4 H+ + O2
-
 Mycobacterium tuberculosis cholest-4-en-3-one-27-oic acid + 4 oxidized ferredoxin [iron-sulfur] cluster + 3 H2O
-
 ?
1.14.15.29 27-hydroxycholest-4-en-3-one + 4 reduced ferredoxin [iron-sulfur] cluster + 4 H+ + O2
-
 Mycobacterium tuberculosis CDC 1551 cholest-4-en-3-one-27-oic acid + 4 oxidized ferredoxin [iron-sulfur] cluster + 3 H2O
-
 ?
1.14.15.29 cholest-4-en-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 physiological substrate, the enzyme hydroxylates cholest-4-en-3-one at C-27 Mycobacterium tuberculosis 27-hydroxycholest-4-en-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.29 cholest-4-en-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 physiological substrate, the enzyme hydroxylates cholest-4-en-3-one at C-27 Mycobacterium tuberculosis CDC 1551 27-hydroxycholest-4-en-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.14.15.29 CYP125A1
-
 Mycobacterium tuberculosis

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.15.29 NADPH
-
 Mycobacterium tuberculosis

General Information

EC Number General Information Comment Organism
1.14.15.29 metabolism CYP125A1 is a key enzyme in cholesterol metabolism and plays a crucial role in circumventing the deleterious effect of cholest-4-en-3-one Mycobacterium tuberculosis
1.14.15.29 physiological function CYP125A1 is required to incorporate the cholesterol side chain carbon atoms into cellular lipids. CYP125A1 is essential for growth of CDC1551 in media containing cholesterol or cholest-4-en-3-one Mycobacterium tuberculosis