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Literature summary for 1.1.3.6 extracted from
- Characterization of cholesterol oxidase from Streptomyces hygroscopicus and Brevibacterium sterolicum (1997), Eur. J. Biochem., 250, 369-376.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| phosphate ions | - |
Streptomyces hygroscopicus |  |
| phosphate ions | activity is enhanced by high concentrations | Brevibacterium sterolicum | |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| propan-2-ol | at 10% propan-2-ol kcat reaches 65% of that at 1.25% propan-2-ol | Streptomyces hygroscopicus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.14 | - |
cholesterol | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Brevibacterium sterolicum | |
| 0.2 | - |
cholesterol | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Streptomyces hygroscopicus | |
| 0.2 | - |
pregnenolone | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Streptomyces hygroscopicus | |
| 0.2 | - |
cholestanol | assay relies on detection of H2O2 formation | Brevibacterium sterolicum | |
| 0.27 | - |
5-Cholesten-3-one | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Brevibacterium sterolicum | |
| 0.3 | - |
3beta-hydroxy-androst-5-en-17-one | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Streptomyces hygroscopicus | |
| 0.4 | - |
pregnenolone | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Brevibacterium sterolicum | |
| 0.5 | - |
3beta-hydroxy-androst-5-en-17-one | assay relies on detection of H2O2 formation | Streptomyces hygroscopicus | |
| 0.7 | - |
cholestanol | assay relies on detection of H2O2 formation | Streptomyces hygroscopicus | |
| 0.8 | - |
3beta-hydroxy-androst-5-en-17-one | assay relies on detection of H2O2 formation | Brevibacterium sterolicum | |
| 1.2 | - |
3beta-hydroxy-androst-5-en-17-one | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Brevibacterium sterolicum | |
| 1.52 | - |
5-Cholesten-3-one | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Streptomyces hygroscopicus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 45000 | - |
gel filtration | Streptomyces hygroscopicus |
| 48000 | - |
gel filtration | Brevibacterium sterolicum |
| 53000 | - |
1 * 53000, SDS-PAGE | Streptomyces hygroscopicus |
| 55000 | - |
1 * 55000, SDS-PAGE | Brevibacterium sterolicum |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| cholesterol + O2 | Streptomyces hygroscopicus | - |
cholest-5-en-3-one + H2O2 | - |
? | |
| cholesterol + O2 | Brevibacterium sterolicum | - |
cholest-5-en-3-one + H2O2 | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Brevibacterium sterolicum | - |
- |
- |
| Streptomyces hygroscopicus | - |
- |
- |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| 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 | |
| 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 | |
| cholesterol + O2 = cholest-5-en-3-one + H2O2 | isomerization is not the rate limiting step | Streptomyces hygroscopicus | |
| cholesterol + O2 = cholest-5-en-3-one + H2O2 | isomerization is not the rate limiting step | Brevibacterium sterolicum |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| cholestanol + O2 | - |
Streptomyces hygroscopicus | ? | - |
? | |
| cholestanol + O2 | - |
Brevibacterium sterolicum | ? | - |
? | |
| cholesterol + O2 | - |
Streptomyces hygroscopicus | cholest-4-en-3-one + H2O2 | - |
? | |
| cholesterol + O2 | - |
Brevibacterium sterolicum | cholest-4-en-3-one + H2O2 | - |
? | |
| cholesterol + O2 | - |
Streptomyces hygroscopicus | cholest-5-en-3-one + H2O2 | - |
? | |
| cholesterol + O2 | - |
Brevibacterium sterolicum | cholest-5-en-3-one + H2O2 | - |
? | |
| pregnenolone + O2 | - |
Streptomyces hygroscopicus | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 55000, SDS-PAGE | Brevibacterium sterolicum |
| monomer | 1 * 53000, SDS-PAGE | Streptomyces hygroscopicus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.8 | - |
cholesterol | assay relies on spectroscopic detection of 4-cholesten-3-one formation, decreasing length of C17 chain affects turnover negatively | Brevibacterium sterolicum | |
| 0.8 | - |
3beta-hydroxy-5-androsten-17-one | assay relies on detection of H2O2 formation | Brevibacterium sterolicum | |
| 7 | - |
3beta-hydroxy-androst-5-en-17-one | assay relies on detection of H2O2 formation | Streptomyces hygroscopicus | |
| 8.2 | - |
3beta-hydroxy-androst-5-en-17-one | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Streptomyces hygroscopicus | |
| 11 | - |
cholesterol | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Streptomyces hygroscopicus | |
| 21 | - |
pregnenolone | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Brevibacterium sterolicum | |
| 24 | - |
pregnenolone | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Streptomyces hygroscopicus | |
| 37 | - |
cholestanol | assay relies on detection of H2O2 formation | Streptomyces hygroscopicus | |
| 40 | - |
cholestanol | assay relies on detection of H2O2 formation | Brevibacterium sterolicum | |
| 67 | - |
cholesterol | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Brevibacterium sterolicum | |
| 278 | - |
5-Cholesten-3-one | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Brevibacterium sterolicum | |
| 332 | - |
5-Cholesten-3-one | assay relies on spectroscopic detection of 4-cholesten-3-one formation | Streptomyces hygroscopicus | |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FAD | - |
Streptomyces hygroscopicus | |
| FAD | non-covalently bound in native enzyme | Brevibacterium sterolicum | |
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