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Literature summary extracted from

  • Pollegioni, L.; Wels, G.; Pilone, M.S.; Ghisla, S.
    Kinetic mechanisms of cholesterol oxidase from Streptomyces hygroscopicus and Brevibacterium sterolicum (1999), Eur. J. Biochem., 264, 140-151.
    View publication on PubMed

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.3.6 0.02
-
cholestanol
-
Brevibacterium sterolicum
1.1.3.6 0.07
-
cholesterol
-
Brevibacterium sterolicum
1.1.3.6 0.11
-
cholesterol
-
Brevibacterium sterolicum
1.1.3.6 0.165
-
O2
-
Brevibacterium sterolicum
1.1.3.6 0.2
-
cholesterol
-
Streptomyces hygroscopicus
1.1.3.6 0.2
-
pregnenolone
-
Streptomyces hygroscopicus
1.1.3.6 0.3
-
3beta-hydroxy-androst-5-en-17-one
-
Streptomyces hygroscopicus
1.1.3.6 0.4
-
pregnenolone
-
Brevibacterium sterolicum
1.1.3.6 0.5
-
3beta-hydroxy-androst-5-en-17-one
-
Streptomyces hygroscopicus
1.1.3.6 0.53
-
O2
-
Streptomyces hygroscopicus
1.1.3.6 0.7
-
cholestanol
-
Streptomyces hygroscopicus
1.1.3.6 0.8
-
cholesterol
-
Streptomyces hygroscopicus
1.1.3.6 0.8
-
3beta-hydroxy-androst-5-en-17-one
-
Brevibacterium sterolicum
1.1.3.6 1.2
-
3beta-hydroxy-androst-5-en-17-one
-
Brevibacterium sterolicum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.3.6 cholesterol + O2 Streptomyces hygroscopicus
-
cholest-5-en-3-one + H2O2
-
?
1.1.3.6 cholesterol + O2 Brevibacterium sterolicum
-
cholest-5-en-3-one + H2O2
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.1.3.6 Brevibacterium sterolicum
-
-
-
1.1.3.6 Streptomyces hygroscopicus
-
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.3.6 cholesterol + O2 = cholest-5-en-3-one + H2O2 bifunctional enzyme, catalyzes both the oxidation of DELTA5-ene-3beta hydroxysteroids with a trans A-B ring junction to the corresponding DELTA5-3-ketosteroid with the reduction of oxygen to hydrogen peroxide, and the isomerization to the DELTA4-3-ketosteroid Streptomyces hygroscopicus
1.1.3.6 cholesterol + O2 = cholest-5-en-3-one + H2O2 bifunctional enzyme, catalyzes both the oxidation of DELTA5-ene-3beta hydroxysteroids with a trans A-B ring junction to the corresponding DELTA5-3-ketosteroid with the reduction of oxygen to hydrogen peroxide, and the isomerization to the DELTA4-3-ketosteroid Brevibacterium sterolicum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.3.6 1-propanol + O2
-
Streptomyces hygroscopicus propanal + H2O2
-
?
1.1.3.6 cholestanol + O2
-
Streptomyces hygroscopicus ?
-
?
1.1.3.6 cholestanol + O2
-
Brevibacterium sterolicum ?
-
?
1.1.3.6 cholesterol + O2 specific for steroid substrates with an OH group in position 3, preference for steroids with at least a C2 chain at position C17 Streptomyces hygroscopicus cholest-4-en-3-one + H2O2
-
?
1.1.3.6 cholesterol + O2 specific for steroid substrates with an OH group in position 3, preference for steroids with at least a C2 chain at position C17 Brevibacterium sterolicum cholest-4-en-3-one + H2O2
-
?
1.1.3.6 cholesterol + O2
-
Streptomyces hygroscopicus cholest-5-en-3-one + H2O2
-
?
1.1.3.6 cholesterol + O2
-
Brevibacterium sterolicum cholest-5-en-3-one + H2O2
-
?
1.1.3.6 cyclohexanol + O2
-
Streptomyces hygroscopicus cyclohexanal + H2O2
-
?
1.1.3.6 epiandrosterone + O2
-
Streptomyces hygroscopicus ?
-
?
1.1.3.6 ethanol + O2
-
Streptomyces hygroscopicus ethanal + H2O2
-
?
1.1.3.6 methanol + O2
-
Streptomyces hygroscopicus methanal + H2O2
-
?
1.1.3.6 pregnenolone + O2
-
Streptomyces hygroscopicus ?
-
?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.3.6 0.8
-
3beta-hydroxy-androst-5-en-17-one product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer Brevibacterium sterolicum
1.1.3.6 0.8
-
3beta-hydroxy-5-androsten-17-one H2O2 production assayed, 100 mM phosphate buffer Brevibacterium sterolicum
1.1.3.6 7
-
epiandrosterone H2O2 production assayed, 100 mM phosphate buffer Streptomyces hygroscopicus
1.1.3.6 8.2
-
3beta-hydroxy-androst-5-en-17-one product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer Streptomyces hygroscopicus
1.1.3.6 21
-
pregnenolone product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer Brevibacterium sterolicum
1.1.3.6 24
-
pregnenolone product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer Streptomyces hygroscopicus
1.1.3.6 28
-
cholesterol 50 mM phosphate buffer, pH 7.5 Streptomyces hygroscopicus
1.1.3.6 37
-
cholestanol H2O2 production assayed, 100 mM phosphate buffer Streptomyces hygroscopicus
1.1.3.6 40
-
cholestanol H2O2 production assayed, 100 mM phosphate buffer Brevibacterium sterolicum
1.1.3.6 48
-
cholesterol 50 mM phosphate buffer, pH 7.5 Brevibacterium sterolicum
1.1.3.6 48.1
-
cholesterol product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer Brevibacterium sterolicum
1.1.3.6 63
-
cholesterol product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer Streptomyces hygroscopicus
1.1.3.6 105
-
cholesterol 50 mM phosphate buffer, pH 7.5, 1% thesit, 10% 2-propanol Brevibacterium sterolicum
1.1.3.6 250
-
5-Cholesten-3-one isomerization to 4-cholesten-3-one, rapid reaction, stopped-flow measurement, 500 mM phosphate buffer, pH 7.5, in the presence of 1% thesit and 1.25% 2-propanol Brevibacterium sterolicum
1.1.3.6 330
-
5-Cholesten-3-one isomerization to 4-cholesten-3-one, rapid reaction, stopped-flow measurement, 500 mM phosphate buffer, pH 7.5, in the presence of 1% thesit and 1.25% 2-propanol Streptomyces hygroscopicus
1.1.3.6 345
-
cholesterol 50 mM phosphate buffer, pH 7.5, 1% thesit, 10% 2-propanol Streptomyces hygroscopicus
1.1.3.6 670
-
5-Cholesten-3-one isomerization to 4-cholesten-3-one, rapid reaction, stopped-flow measurement, 50 mM phosphate buffer, pH 7.5, in the presence of 1% thesit and 1.25% 2-propanol Streptomyces hygroscopicus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.3.6 FAD non-covalently bound Streptomyces hygroscopicus
1.1.3.6 FAD covalently bound in His-tagged recombinant enzyme Brevibacterium sterolicum