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1.1.3.6: cholesterol oxidase

This is an abbreviated version!
For detailed information about cholesterol oxidase, go to the full flat file.

Word Map on EC 1.1.3.6

Reaction

cholesterol
+
O2
=
cholest-5-en-3-one
 +
H2O2

Synonyms

3beta-hydroxy steroid oxidoreductase, 3beta-hydroxysteroid: oxygen oxidoreductase, 3beta-hydroxysteroid:oxygen oxidoreductase, 3beta-hydroxysterol oxidase, BsChOx, CgChoA, CHO, CHO-U, ChO3, ChoA, choBb, CHOD, ChoG, ChoL, cholesterol oxidase, cholesterol oxidase I, cholesterol oxidase II, cholesterol-O2 oxidoreductase, CHOLOX, choM, ChoM1, ChoM2, choP, ChoRI, ChoRII, ChoS, ChOx, CO, CO1, COase, COD, COD-B, COX, HCEO-forming enzyme, HMPREF0204_11499, oxidase, cholesterol, PimE, ShChOx, type I ChOx

ECTree

     1 Oxidoreductases
         1.1 Acting on the CH-OH group of donors
             1.1.3 With oxygen as acceptor
                1.1.3.6 cholesterol oxidase

Purification

Purification on EC 1.1.3.6 - cholesterol oxidase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate, DEAE-cellulose, butyl-toyopearl
-
ammonium sulfate, ion-exchange chromatography, gel filtration
-
butyl-Sepharose column chromatography and DEAE-cellulose column chromatography
choleterylglycine-CM-cellulose
-
column nickel affinity chromatography
DEAE-cellulose, cholesterol affinity column, Sephadex G-150
-
DEAE-Sepharose column chromatography
effective method for the preparation of active PsChO3 using urea as denaturing agent, combined with his-tag trapping and on-column refolding. Using this method, the enzyme is successfully refolded from inclusion bodies expressed in Escherichia coli
extracellular enzyme 59fold from strain PKPD-CL by ammonium sulfate fractionation, anion exchange chromatography, ultrafiltration, and gel filtration
-
HiTrap Ni-chelating column chromatography
-
inducible and constitutive enzyme
-
native enzyme 2.3fold by ammnium sulfate fractionation and gel filtration
-
native extracellular enzyme 14.3fold by ammonium sulfate fractionation and affinity chromatography
-
native extracellular enzyme 18fold by anion exchange chromatography and gel filtration
-
native extracellular enzyme 28.4fold from cell culture supernatant by ammonium sulfate fractionation, dialysis, and riboflavin affinity chromatography
-
native extracellular enzyme from medium 12fold to homogeneity by ammonium sulfate fractionation, dialysis, ultrafiltration, anion exchange chromatography and gel filtration
-
Ni2+-chelating Sepharose column chromatography
overview: purification procedures
Nocardia erythropolis
-
partially purified by precipitation with ammonium sulfate
-
polyethylene glycol 4000 precipitation, diethylaminoethyl Sephacel anionic column chromatography and Superdex-200 gel filtration
-
recombinant His-tagged isozyme CHOM1 from Bacillus subtilis strain 168/pMA5 by nickel affinity chromatography
recombinant His-tagged isozyme CHOM2 from Bacillus subtilis strain 168/pMA5 by nickel affinity chromatography
recombinant His6-tagged enzyme from Escherichia coli train BL21(DE3) by nickel affinity chromatography, dialysis and ultrafiltration
recombinant N-terminally His-tagged enzyme from Escherichia coli strain JM109 by nickel affinity chromatography
recombinant wild-type and mutant enzymes with with His-tags both at the N-terminus and at the C-terminus from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography
-
salting out and ion-exchange chromatography
-
strain ST-200, ammonium sulfate, DEAE-cellulose, butyl-toyopearl, Sephadex G-100
-
to purify the recombinant enzyme, 500 mg cholesterol is added to 50 ml of crude extract, and then incubated for 2 h at 4°C followed by centrifugation at 10000g for 10 min
wild-type and mutant enzymes
-
wild-type and mutant enzymes V145Q, V145E, S379A and S379V
-