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Literature summary for 1.1.3.6 extracted from
- Cloning, expression and biochemical characterization of the cholesterol oxidase CgChoA from Chryseobacterium gleum (2014), BMC Biotechnol., 14, 46 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene HMPREF0204_11499, phylogenetic tree, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain JM109. Only when the cultivation temperature is decreased to 16°C immediately after induction, soluble and active protein is present | Chryseobacterium gleum |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics | Chryseobacterium gleum | |
| 0.5 | - |
taurocholate | pH 6.75, 35°C | Chryseobacterium gleum |  |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| intracellular | the lack of a signal peptide indicated the intracellular localization of the enzyme in the native host | Chryseobacterium gleum | 5622 | - |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 85000 | - |
recombinant N-terminally His-tagged enzyme, gel filtration | Chryseobacterium gleum |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Chryseobacterium gleum | cholesterol oxidase is a bifunctional enzyme that catalyzes the oxidation and isomerization of cholesterol to cholest-4-en-3-one while dioxygen is finally reduced to H2O2 as by-product. The enzymatic overall cholesterol oxidation comprises three steps. In the first one the 3beta-OH group of cholesterol is oxidized to the corresponding ketone with the concomitant reduction of the FAD cofactor. In a second step an isomerization of the double bond from the DELTA5-6 position to the DELTA4-5 position takes place. The FAD is recycled in a redox reaction with dioxygen, yielding hydrogen peroxide | ? | - |
? | |
| additional information | Chryseobacterium gleum DSM 16776 | cholesterol oxidase is a bifunctional enzyme that catalyzes the oxidation and isomerization of cholesterol to cholest-4-en-3-one while dioxygen is finally reduced to H2O2 as by-product. The enzymatic overall cholesterol oxidation comprises three steps. In the first one the 3beta-OH group of cholesterol is oxidized to the corresponding ketone with the concomitant reduction of the FAD cofactor. In a second step an isomerization of the double bond from the DELTA5-6 position to the DELTA4-5 position takes place. The FAD is recycled in a redox reaction with dioxygen, yielding hydrogen peroxide | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Chryseobacterium gleum | A0A3S4PHH6 | - |
- |
| Chryseobacterium gleum DSM 16776 | A0A3S4PHH6 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant N-terminally His-tagged enzyme from Escherichia coli strain JM109 by nickel affinity chromatography | Chryseobacterium gleum |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 15.5 | - |
purified His-tagged enzyme, pH 6.75, 35°C | Chryseobacterium gleum |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| cholesterol + O2 | - |
Chryseobacterium gleum | cholest-4-en-3-one + H2O2 | - |
? | |
| cholesterol + O2 | - |
Chryseobacterium gleum DSM 16776 | cholest-4-en-3-one + H2O2 | - |
? | |
| additional information | cholesterol oxidase is a bifunctional enzyme that catalyzes the oxidation and isomerization of cholesterol to cholest-4-en-3-one while dioxygen is finally reduced to H2O2 as by-product. The enzymatic overall cholesterol oxidation comprises three steps. In the first one the 3beta-OH group of cholesterol is oxidized to the corresponding ketone with the concomitant reduction of the FAD cofactor. In a second step an isomerization of the double bond from the DELTA5-6 position to the DELTA4-5 position takes place. The FAD is recycled in a redox reaction with dioxygen, yielding hydrogen peroxide | Chryseobacterium gleum | ? | - |
? | |
| additional information | the enzyme also acts as a DELTA5-ketosteroid isomerase, EC 5.3.3.1, converting a 3-oxo-DELTA5-steroid to a 3-oxo-DELTA4-steroid | Chryseobacterium gleum | ? | - |
? | |
| additional information | cholesterol oxidase is a bifunctional enzyme that catalyzes the oxidation and isomerization of cholesterol to cholest-4-en-3-one while dioxygen is finally reduced to H2O2 as by-product. The enzymatic overall cholesterol oxidation comprises three steps. In the first one the 3beta-OH group of cholesterol is oxidized to the corresponding ketone with the concomitant reduction of the FAD cofactor. In a second step an isomerization of the double bond from the DELTA5-6 position to the DELTA4-5 position takes place. The FAD is recycled in a redox reaction with dioxygen, yielding hydrogen peroxide | Chryseobacterium gleum DSM 16776 | ? | - |
? | |
| additional information | the enzyme also acts as a DELTA5-ketosteroid isomerase, EC 5.3.3.1, converting a 3-oxo-DELTA5-steroid to a 3-oxo-DELTA4-steroid | Chryseobacterium gleum DSM 16776 | ? | - |
? | |
| taurocholate + O2 | - |
Chryseobacterium gleum | ? + H2O2 | - |
? | |
| taurocholate + O2 | - |
Chryseobacterium gleum DSM 16776 | ? + H2O2 | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 x 60400, about, recombinant N-terminally His-tagged enzyme, sequence calculation, 1 * 60000, recombinant N-terminally His-tagged enzyme, SDS-PAGE | Chryseobacterium gleum |
| More | the overall enzyme structure comprises two domains, the FAD binding domain and the substrate binding domain, structure comparisons | Chryseobacterium gleum |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CgChoA | - |
Chryseobacterium gleum |
| ChoA | - |
Chryseobacterium gleum |
| HMPREF0204_11499 | - |
Chryseobacterium gleum |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 35 | - |
- |
Chryseobacterium gleum |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.75 | - |
- |
Chryseobacterium gleum |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FAD | dependent on, the enzyme contains a Rossmann fold (xh)2GxGxxGx(xxh)2(x) FAD binding site, where x is any amino acid and h an hydrophobic one, between V44 and E70 in the N-terminal region. CgChoA belongs to the non-covalent FAD-dependent enzymes belonging to the class I family. Residues N503 and Y464 are required for stabilization of the reduced form cofactor-enzyme binding | Chryseobacterium gleum | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | CgChoA belongs to the non-covalent FAD-dependent enzymes belonging to the class I family | Chryseobacterium gleum |
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