Literature summary extracted from
Golden, E.; Karton, A.; Vrielink, A.
High-resolution structures of cholesterol oxidase in the reduced state provide insights into redox stabilization (2014), Acta Crystallogr. Sect. D, 70, 3155-3166 .
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
1.1.3.6 |
recombinant expression of His6-tagged enzyme in Escherichia coli train BL21(DE3) |
Streptomyces sp. SA-COO |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
1.1.3.6 |
purified nontagged and His6-tagged enzyme reduced with dithionite under aerobic conditions and in the presence of the substrate 2-propanol under both aerobic and anaerobic conditions, vapour diffusion using the hanging-drop method, mixing of 7 mg/ml protein in 50 mM HEPES pH 7.0 with reservoir solution containing 7% PEG 8000, 100 mM cacodylate, pH 5.2, 125 mM MnSO4, followed by microseeding in 12% PEG 8000, 100 mM sodium cacodylate, pH 5.2, 125 mM MnSO4, X-ray diffraction structure determination and analysis at 1.12-1.34 A resolution |
Streptomyces sp. SA-COO |
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 sp. SA-COO |
via cholest-5-en-3-one |
cholest-4-en-3-one + H2O2 |
- |
? |
|
1.1.3.6 |
additional information |
Streptomyces sp. SA-COO |
cholesterol oxidase is a bifunctional flavoenzyme that catalyzes the oxidation and isomerization of cholesterol to cholest-4-en-3-one |
? |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
1.1.3.6 |
Streptomyces sp. SA-COO |
P12676 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
1.1.3.6 |
recombinant His6-tagged enzyme from Escherichia coli train BL21(DE3) by nickel affinity chromatography, dialysis and ultrafiltration |
Streptomyces sp. SA-COO |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
1.1.3.6 |
cholesterol + O2 |
via cholest-5-en-3-one |
Streptomyces sp. SA-COO |
cholest-4-en-3-one + H2O2 |
- |
? |
|
1.1.3.6 |
additional information |
cholesterol oxidase is a bifunctional flavoenzyme that catalyzes the oxidation and isomerization of cholesterol to cholest-4-en-3-one |
Streptomyces sp. SA-COO |
? |
- |
? |
|
1.1.3.6 |
additional information |
the enzyme catalyzes the oxidation and isomerization of steroids containing a 3-hydroxyl group, with the cholestane group conferring binding specificity in the active site. The enzyme is also able to oxidize small-molecule alcohols such as 2-propanol and methanol, with a preference for those containing aromatic ring, but with lower activity. Substrate oxidation results in reduction of the cofactor during the reductive half reaction. An oxidative half reaction occurs with re-oxidation of the flavin cofactor by molecular oxygen to form hydrogen peroxide. Structure-function relationship and analysis overview |
Streptomyces sp. SA-COO |
? |
- |
? |
|
Cofactor
EC Number |
Cofactor |
Comment |
Organism |
Structure |
---|
1.1.3.6 |
FAD |
the structure of the dithionite-reduced enzyme reveals a sulfite molecule covalently bound to the FAD cofactor. The hydride transfer generates a tetrahedral geometry about the flavin N5 atom |
Streptomyces sp. SA-COO |
|
General Information
EC Number |
General Information |
Comment |
Organism |
---|
1.1.3.6 |
evolution |
the type I CO from Streptomyces sp. SA-COO is a member of the glucosemethanolcholine (GMC) oxidoreductase family and contains a single molecule of flavin adenine dinucleotide (FAD) noncovalently but tightly bound to the protein |
Streptomyces sp. SA-COO |
1.1.3.6 |
additional information |
the structure of the dithionite-reduced enzyme reveals a sulfite molecule covalently bound to the FAD cofactor. The isoalloxazine ring system displays a bent structure relative to that of the oxidized enzyme, and alternate conformations of a triad of aromatic residues near to the cofactor are evident. The strutcure of anaerobically trapped reduced enzyme structure in the presence of 2-propanol does not show a similar bending of the flavin ring system, but does show alternate conformations of the aromatic triad. The hydride transfer generates a tetrahedral geometry about the flavin N5 atom |
Streptomyces sp. SA-COO |