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Literature summary extracted from

  • Qin, H.M.; Zhu, Z.; Ma, Z.; Xu, P.; Guo, Q.; Li, S.; Wang, J.W.; Mao, S.; Liu, F.; Lu, F.
    Rational design of cholesterol oxidase for efficient bioresolution of cholestane skeleton substrates (2017), Sci. Rep., 7, 16375 .
    View publication on PubMedView publication on EuropePMC

Protein Variants

EC Number Protein Variants Comment Organism
1.1.3.6 F70A improved activities toward cholestane skeleton substrates. Improved activity toward beta-sitosterol, dehydroepiandrosterone, pregnenolone and stigmasterol Pimelobacter simplex
1.1.3.6 F70L the mutant enzyme shows almost the same level of activity as the wild-type enzyme Pimelobacter simplex
1.1.3.6 F70V mutation increases the binding free energy between the mutant enzyme (PsChO) and cholesterol and might cause the movement of loops L56-P77, K45-P49 and L350-E354 at active site. Improved activities toward cholestane skeleton substrates. 50% higher activities toward cholesterol when compared with that of the wild-type enzyme Pimelobacter simplex
1.1.3.6 V64C mutation increases the binding free energy between the mutant enzyme (PsChO) and cholesterol and might cause the movement of loops L56-P77, K45-P49 and L350-E354 at active site. Improved activities toward cholestane skeleton substrates. 50% higher activities toward cholesterol when compared with that of the wild-type enzyme Pimelobacter simplex
1.1.3.6 V64I improved activities toward cholestane skeleton substrates. Improved activity toward beta-sitosterol, dehydroepiandrosterone, pregnenolone and stigmasterol Pimelobacter simplex
1.1.3.6 V64L improved activities toward cholestane skeleton substrates. Improved activity toward beta-sitosterol, dehydroepiandrosterone, pregnenolone and stigmasterol Pimelobacter simplex

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.3.6 0.15483
-
 cholesterol pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 0.17831
-
 cholesterol pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 0.17842
-
 beta-sitosterol pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 0.19061
-
 pregnenolone pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 0.20485
-
 cholesterol pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 0.22076
-
 beta-sitosterol pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 0.22842
-
 pregnenolone pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 0.27352
-
 dehydroepiandrosterone pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 0.29455
-
 pregnenolone pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 0.31881
-
 dehydroepiandrosterone pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 0.32861
-
 beta-sitosterol pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 0.34121
-
 dehydroepiandrosterone pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 0.38412
-
 stigmasterol pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 0.52884
-
 stigmasterol pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 0.78961
-
 stigmasterol pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex

Organism

EC Number Organism UniProt Comment Textmining
1.1.3.6 Pimelobacter simplex
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.3.6 beta-sitosterol + O2
-
 Pimelobacter simplex ? + H2O2
-
 ?
1.1.3.6 cholesterol + O2
-
 Pimelobacter simplex cholest-5-en-3-one + H2O2
-
 ?
1.1.3.6 dehydroepiandrosterone + O2
-
 Pimelobacter simplex androst-5-en-3,17-dione + H2O2
-
 ?
1.1.3.6 pregnenolone + O2
-
 Pimelobacter simplex 5-pregnen-3,20-dione + H2O2
-
 ?
1.1.3.6 stigmasterol + O2
-
 Pimelobacter simplex stigmasta-5,22-dien-3-one + H2O2
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.3.6 25
-
-
 Pimelobacter simplex

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.3.6 50
-
 30 min, 25% residual activity Pimelobacter simplex

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.3.6 0.08
-
 stigmasterol pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 1.58
-
 stigmasterol pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 4.23
-
 stigmasterol pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 6.82
-
 dehydroepiandrosterone pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 9.56
-
 dehydroepiandrosterone pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 9.86
-
 beta-sitosterol pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 11.78
-
 pregnenolone pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 12.29
-
 cholesterol pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 13.25
-
 beta-sitosterol pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 13.68
-
 dehydroepiandrosterone pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 13.71
-
 pregnenolone pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 15.25
-
 pregnenolone pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 15.48
-
 cholesterol pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 16.06
-
 beta-sitosterol pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 23.02
-
 cholesterol pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.3.6 7.5
-
-
 Pimelobacter simplex

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.3.6 5 9 more than 75% of the maximal activity is retained over the pH range of 5.0-9.0 Pimelobacter simplex

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.1.3.6 5.5 9 good pH stability over the pH range of 5.0-9.0 Pimelobacter simplex

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.3.6 2
-
 stigmasterol pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 8
-
 stigmasterol pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 20
-
 dehydroepiandrosterone pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 20
-
 stigmasterol pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 30
-
 beta-sitosterol pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 30
-
 dehydroepiandrosterone pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 40
-
 pregnenolone pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 50
-
 dehydroepiandrosterone pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 60
-
 cholesterol pH 7.5, temperature not specified in the publication, wild-type enzyme Pimelobacter simplex
1.1.3.6 60
-
 pregnenolone pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 60
-
 beta-sitosterol pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex
1.1.3.6 80
-
 pregnenolone pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 90
-
 beta-sitosterol pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 100
-
 cholesterol pH 7.5, temperature not specified in the publication, mutant enzyme F70V Pimelobacter simplex
1.1.3.6 130
-
 cholesterol pH 7.5, temperature not specified in the publication, mutant enzyme V64C Pimelobacter simplex