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Literature summary for 1.1.3.6 extracted from

  • Doukyu, N.; Nihei, S.
    Cholesterol oxidase with high catalytic activity from Pseudomonas aeruginosa Screening, molecular genetic analysis, expression and characterization (2015), J. Biosci. Bioeng., 120, 24-30 .
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
Sarcosyl activates at 0.5% w/v Pseudomonas aeruginosa
sodium cholate activates at 0.5% w/v Pseudomonas aeruginosa
Toluene 12% activation in a ration of 1:2 Pseudomonas aeruginosa
Triton X-100 activates at 0.5% w/v Pseudomonas aeruginosa
Triton X-405 activates at 0.5% w/v Pseudomonas aeruginosa
Tween 20 activates at 0.5% w/v Pseudomonas aeruginosa

Cloned(Commentary)

Cloned (Comment) Organism
gene choP, DNA and amino acid sequence determination and analysis, sequence comparison and phylogenetic tree, subcloning in Escherichia coli strain DH5alpha, recombinant overexpression in Escherichia coli strain Rosetta (DE3) pLysS in inclusion bodies from cells grown at 20°C and 30°C. A soluble and active PA157 enzyme is obtained when the recombinant cells were grown at 10°C Pseudomonas aeruginosa

Inhibitors

Inhibitors Comment Organism Structure
SDS complete inhibition Pseudomonas aeruginosa
sodium laurylbenzenesulfonate complete inhibition Pseudomonas aeruginosa
sodium polyoxyethylene alkyl ether sulfate
-
Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0926
-
cholesterol pH 7.0-8.0, 30°C Pseudomonas aeruginosa

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular
-
Pseudomonas aeruginosa
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
additional information 1 mM of Ca2+, Mg2+, Mn2+, Ni2+, Fe2+, and Zn2+ have no obvious effect on the enzyme activity.No effect by EDTA Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
cholesterol + O2 Pseudomonas aeruginosa
-
6beta-hydroxycholest-4-en-3-one + H2O2
-
?
cholesterol + O2 Pseudomonas aeruginosa PA157
-
6beta-hydroxycholest-4-en-3-one + H2O2
-
?

Organic Solvent Stability

Organic Solvent Comment Organism
Acetone in a ratio 1:2 with enzyme solution, pH 7.0, 37°C, 24 h, 2% activity remaining Pseudomonas aeruginosa
chloroform in a ratio 1:2 with enzyme solution, pH 7.0, 37°C, 24 h, 89% activity remaining Pseudomonas aeruginosa
cyclooctane in a ratio 1:2 with enzyme solution, pH 7.0, 37°C, 24 h, 108% activity remaining Pseudomonas aeruginosa
Ethanol in a ratio 1:2 with enzyme solution, pH 7.0, 37°C, 24 h, 31% activity remaining Pseudomonas aeruginosa
isopropanol in a ratio 1:2 with enzyme solution, pH 7.0, 37°C, 24 h, 6% activity remaining Pseudomonas aeruginosa
Methanol in a ratio 1:2 with enzyme solution, pH 7.0, 37°C, 24 h, 65% activity remaining Pseudomonas aeruginosa
p-xylene in a ratio 1:2 with enzyme solution, pH 7.0, 37°C, 24 h, 102% activity remaining Pseudomonas aeruginosa
toluene in a ratio 1:2 with enzyme solution, pH 7.0, 37°C, 24 h, 112% activity remaining Pseudomonas aeruginosa

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa X5IDD2
-
-
Pseudomonas aeruginosa PA157 X5IDD2
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-betacholestanol + O2
-
Pseudomonas aeruginosa ? + H2O2
-
?
3-betacholestanol + O2
-
Pseudomonas aeruginosa PA157 ? + H2O2
-
?
beta-sitosterol + O2
-
Pseudomonas aeruginosa stigmast-5-en-3-one + H2O2
-
?
beta-sitosterol + O2
-
Pseudomonas aeruginosa PA157 stigmast-5-en-3-one + H2O2
-
?
beta-stigmasterol + O2
-
Pseudomonas aeruginosa stigmasta-5,22-dien-3-one + H2O2
-
?
beta-stigmasterol + O2
-
Pseudomonas aeruginosa PA157 stigmasta-5,22-dien-3-one + H2O2
-
?
cholesterol + O2
-
Pseudomonas aeruginosa 6beta-hydroxycholest-4-en-3-one + H2O2
-
?
cholesterol + O2
-
Pseudomonas aeruginosa PA157 6beta-hydroxycholest-4-en-3-one + H2O2
-
?
additional information the enzyme oxidizes 3beta-hydroxysteroids such as cholesterol, beta-cholestanol, and beta-sitosterol at high rates. No activity with 5alpha-androstan-3beta-ol-17-one, and 3beta-hydroxyandrost-5-en-17-one Pseudomonas aeruginosa ?
-
?
additional information the enzyme oxidizes 3beta-hydroxysteroids such as cholesterol, beta-cholestanol, and beta-sitosterol at high rates. No activity with 5alpha-androstan-3beta-ol-17-one, and 3beta-hydroxyandrost-5-en-17-one Pseudomonas aeruginosa PA157 ?
-
?
pregnenolone + O2 low activity Pseudomonas aeruginosa pregn-5-ene-3,20-dione + H2O2
-
?

Subunits

Subunits Comment Organism
? x * 65344, mass spectrometric analysis Pseudomonas aeruginosa

Synonyms

Synonyms Comment Organism
choP
-
Pseudomonas aeruginosa
HCEO-forming enzyme
-
Pseudomonas aeruginosa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
-
Pseudomonas aeruginosa

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4 65 purified recombinant enzyme, pH 7.0, 30 min, stable at Pseudomonas aeruginosa
70
-
purified recombinant enzyme, pH 7.0, 30 min, 90% activity remaining Pseudomonas aeruginosa
75 80 purified recombinant enzyme, pH 7.0, 30 min, inactivation Pseudomonas aeruginosa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
Pseudomonas aeruginosa

pH Stability

pH Stability pH Stability Maximum Comment Organism
1 2 purified recombinant enzyme, 30°C, 10% activity remaining after 30 min Pseudomonas aeruginosa
5.5 10 purified recombinant enzyme, 30°C, 30 min, stable at Pseudomonas aeruginosa
11
-
purified recombinant enzyme, 30°C, 30 min, 70% activity remaining Pseudomonas aeruginosa

Cofactor

Cofactor Comment Organism Structure
FAD
-
Pseudomonas aeruginosa

pI Value

Organism Comment pI Value Maximum pI Value
Pseudomonas aeruginosa isoelectric focusing
-
5.96