Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.3.6 extracted from

  • Lyubimov, A.Y.; Chen, L.; Sampson, N.S.; Vrielink, A.
    A hydrogen-bonding network is important for oxidation and isomerization in the reaction catalyzed by cholesterol oxidase (2009), Acta Crystallogr. Sect. D, 65, 1222-1231.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3)pLysS cells Streptomyces sp.

Crystallization (Commentary)

Crystallization (Comment) Organism
mutant enzymes N485D and N485L, hanging drop vapor diffusion method, using 9-11% (w/v) polyethylene glycol 8000, 75 mM MnSO4 and 100 mM sodium cacodylate pH 5.2 Streptomyces sp.

Protein Variants

Protein Variants Comment Organism
N485D the kcat of N485D is diminished about 650times compared with that of wild type, while the apparent Km value is minimally affected. For the N485D mutant, pH 5.1results in a 20fold increase in the rate of oxidation compared with that at pH 7.0 Streptomyces sp.
N485L the kcat of N485D is diminished about 1110times compared with that of wild type, while the apparent Km value is minimally affected Streptomyces sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0027
-
cholesterol apparent value, wild type enzyme, pH 7.0, temperature not specified in the publication Streptomyces sp.
0.0046
-
cholesterol apparent value, mutant enzyme N485L, pH 7.0, temperature not specified in the publication Streptomyces sp.
0.0062
-
cholesterol apparent value, mutant enzyme N485L, pH 5.1, temperature not specified in the publication Streptomyces sp.
0.0063
-
cholesterol apparent value, mutant enzyme N485D, pH 5.1, temperature not specified in the publication Streptomyces sp.
0.0066
-
cholesterol apparent value, mutant enzyme N485D, pH 7.0, temperature not specified in the publication Streptomyces sp.
0.0067
-
cholesterol apparent value, wild type enzyme, pH 5.1, temperature not specified in the publication Streptomyces sp.

Organism

Organism UniProt Comment Textmining
Streptomyces sp. P12676
-
-

Purification (Commentary)

Purification (Comment) Organism
butyl-Sepharose column chromatography and DEAE-cellulose column chromatography Streptomyces sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cholesterol + O2
-
Streptomyces sp. cholest-4-en-3-one + H2O2
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.035
-
cholesterol apparent value, mutant enzyme N485L, pH 5.1, temperature not specified in the publication Streptomyces sp.
0.044
-
cholesterol apparent value, mutant enzyme N485L, pH 7.0, temperature not specified in the publication Streptomyces sp.
0.073
-
cholesterol apparent value, mutant enzyme N485D, pH 7.0, temperature not specified in the publication Streptomyces sp.
1.4
-
cholesterol apparent value, mutant enzyme N485D, pH 5.1, temperature not specified in the publication Streptomyces sp.
46
-
cholesterol apparent value, wild type enzyme, pH 5.1, temperature not specified in the publication Streptomyces sp.
47
-
cholesterol apparent value, wild type enzyme, pH 7.0, temperature not specified in the publication Streptomyces sp.

Cofactor

Cofactor Comment Organism Structure
FAD
-
Streptomyces sp.