Information on EC 1.1.3.6 - cholesterol oxidase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY
1.1.3.6
-
RECOMMENDED NAME
GeneOntology No.
cholesterol oxidase
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
oxidation
-
-
oxidation
-
-
-
redox reaction
-
-
radical or hydride transfer mechanism
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
cholesterol degradation to androstenedione I (cholesterol oxidase)
-
Steroid degradation
-
SYSTEMATIC NAME
IUBMB Comments
cholesterol:oxygen oxidoreductase
Contains FAD. Cholesterol oxidases are secreted bacterial bifunctional enzymes that catalyse the first two steps in the degradation of cholesterol. The enzyme catalyses the oxidation of the 3beta-hydroxyl group to a keto group, and the isomerization of the double bond in the oxidized steroid ring system from the Delta5 position to Delta6 position (cf. EC 5.3.3.1, steroid Delta-isomerase).
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3beta-hydroxy steroid oxidoreductase
-
-
-
-
3beta-hydroxysteroid:oxygen oxidoreductase
-
-
-
-
3beta-hydroxysterol oxidase
-
-
3beta-hydroxysterol oxidase
Mycobacterium sp., Nocardia erythropolis, Nocardia rhodochrous, Pseudomonas spp.
-
-
3beta-hydroxysterol oxidase
-
-
3beta-hydroxysterol oxidase
-
-
3beta-hydroxysterol oxidase
-
-
3beta-hydroxysterol oxidase
-
-
3beta-hydroxysterol oxidase
Streptomyces violascens, Streptoverticillum cholesterolicum, uncultured gamma proteobacterium
-
-
3beta-hydroxysterol oxidase
uncultured gamma proteobacterium Y-134
-
-
-
CHO
H9NJ43, H9NJ52
-
CHO
Streptomyces parvus 13647 J
-
-
-
choBb
Q2I2N2
-
choBb
Brevibacterium sp. CCTCC M201008
Q2I2N2
-
-
CHOD
-
-
-
-
ChoG
C5II17
a class II cholesterol oxidase
ChoG
Rhodococcus sp. CECT3014
C5II17
a class II cholesterol oxidase
-
ChoL
A7LGL0
a fragment of a cholesterol oxidase
ChoL
Streptomyces virginiae IBL-14
A7LGL0
a fragment of a cholesterol oxidase
-
cholesterol oxidase
-
-
cholesterol oxidase
-
-
cholesterol oxidase
Mycobacterium sp., Nocardia erythropolis, Nocardia rhodochrous, Pseudomonas spp., Rhodococcus equi, Schizophyllum commune
-
-
cholesterol oxidase
-
-
cholesterol oxidase
Streptomyces violascens, Streptoverticillum cholesterolicum, uncultured gamma proteobacterium
-
-
cholesterol oxidase
uncultured gamma proteobacterium Y-134
-
-
-
cholesterol oxidase I
-
-
cholesterol oxidase II
-
-
cholesterol-O2 oxidoreductase
-
-
-
-
CHOLOX
B5MGF8
belongs to the type II cholesterol oxidases
ChoM2
H9NJ43, H9NJ52
-
ChOx
Mycobacterium sp., Nocardia erythropolis, Nocardia rhodochrous
-
-
ChOx
-
-
ChOx
Streptomyces violascens, Streptoverticillum cholesterolicum, uncultured gamma proteobacterium
-
-
ChOx
uncultured gamma proteobacterium Y-134
-
-
-
CO1
Bacillus sp. SFF34
-
-
-
COD
-
-
-
COD
Brevibacterium sp. M201008
-
-
-
COD
Streptomyces sp. R-6
-
-
-
oxidase, cholesterol
-
-
-
-
type I ChOx
-
-
CAS REGISTRY NUMBER
COMMENTARY
9028-76-6
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Arthrobacter sp. IM79
-
-
-
Manually annotated by BRENDA team
SFF34
-
-
Manually annotated by BRENDA team
Bacillus sp. SFF34
SFF34
-
-
Manually annotated by BRENDA team
isolated from a soil sample collected from the riverside of the Yellow River in Jinan City, Shandong Province, China
-
-
Manually annotated by BRENDA team
isolated from a soil sample collected from the riverside of the Yellow River in Jinan City, Shandong Province, China
-
-
Manually annotated by BRENDA team
CCTCC M201008
SwissProt
Manually annotated by BRENDA team
Brevibacterium sp. CCTCC M201008
CCTCC M201008
SwissProt
Manually annotated by BRENDA team
Brevibacterium sp. M201008
-
-
-
Manually annotated by BRENDA team
nov. sp. ATCC21387
-
-
Manually annotated by BRENDA team
strain ST-200
-
-
Manually annotated by BRENDA team
Burkholderia cepacia ST-200
-
UniProt
Manually annotated by BRENDA team
Burkholderia cepacia ST-200
strain ST-200
-
-
Manually annotated by BRENDA team
-
Q13UN0
UniProt
Manually annotated by BRENDA team
Corynebacterium cholesterolicum
-
-
-
Manually annotated by BRENDA team
Corynebacterium urealyticum DSM7109
-
UniProt
Manually annotated by BRENDA team
isozyme ChoM1
UniProt
Manually annotated by BRENDA team
isozyme ChoM2; several strains
UniProt
Manually annotated by BRENDA team
probably identical with cholesterol decompositing Mycobacterium cholesterolicum
-
-
Manually annotated by BRENDA team
Nocardia farcinica IFM10152
-
UniProt
Manually annotated by BRENDA team
Nocardia rhodochrous
-
-
-
Manually annotated by BRENDA team
Nocardia rhodochrous
3 forms
-
-
Manually annotated by BRENDA team
Nocardia rhodochrous
renamed as Rhodococcus rhodochrous
-
-
Manually annotated by BRENDA team
strain F2, gene choF
SwissProt
Manually annotated by BRENDA team
Pimelobacter simplex F2
strain F2, gene choF
SwissProt
Manually annotated by BRENDA team
ST-200
-
-
Manually annotated by BRENDA team
Pseudomonas sp. COX629
-
-
-
Manually annotated by BRENDA team
Pseudomonas sp. ST-200
ST-200
-
-
Manually annotated by BRENDA team
Pseudomonas spp.
-
-
-
Manually annotated by BRENDA team
high levels of ChOx activity, significant similarities to putative ChOxs encoded by Mycobacterium tuberculosis and Mycobacterium leprae
-
-
Manually annotated by BRENDA team
Rhodococcus equi ATCC33706
-
UniProt
Manually annotated by BRENDA team
ATCC 25544, de novo enzyme synthesis is induced by cholesterol
-
-
Manually annotated by BRENDA team
solated from soil
-
-
Manually annotated by BRENDA team
Rhodococcus sp. CECT3014
-
UniProt
Manually annotated by BRENDA team
Rhodococcus sp. GK1
solated from soil
-
-
Manually annotated by BRENDA team
fragment
SwissProt
Manually annotated by BRENDA team
from soil sample collected from effluent sediment of a steel plant in Barabanki (Uttar Pradesh, India)
-
-
Manually annotated by BRENDA team
Streptomyces parvus 13647 J
from soil sample collected from effluent sediment of a steel plant in Barabanki (Uttar Pradesh, India)
-
-
Manually annotated by BRENDA team
strain A19249
-
-
Manually annotated by BRENDA team
strain SA-COO
-
-
Manually annotated by BRENDA team
Streptomyces sp. A19249
strain A19249
-
-
Manually annotated by BRENDA team
Streptomyces sp. R-6
-
-
-
Manually annotated by BRENDA team
strain SA-COO
-
-
Manually annotated by BRENDA team
Streptomyces virginiae IBL-14
-
UniProt
Manually annotated by BRENDA team
Streptoverticillium cholesterolieum
-
-
-
Manually annotated by BRENDA team
Streptoverticillum cholesterolicum
-
-
-
Manually annotated by BRENDA team
uncultured gamma proteobacterium Y-134
-
-
-
Manually annotated by BRENDA team
uncultured gamma proteobacterium Y-134
Y-134
-
-
Manually annotated by BRENDA team
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.8
-
3beta-hydroxy-5-androsten-17-one
-
assay relies on detection of H2O2 formation
0.8
-
3beta-hydroxy-5-androsten-17-one
-
H2O2 production assayed, 100 mM phosphate buffer
0.8
-
3beta-hydroxy-androst-5-en-17-one
-
product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer
7
-
3beta-hydroxy-androst-5-en-17-one
-
assay relies on detection of H2O2 formation
8.2
-
3beta-hydroxy-androst-5-en-17-one
-
assay relies on spectroscopic detection of 4-cholesten-3-one formation
8.2
-
3beta-hydroxy-androst-5-en-17-one
-
product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer
28
-
5-Cholesten-3-one
-
H69A mutant enzyme, 5-cholesten-3-one isomerization, 50 mM phosphate buffer, 1% thesit, 1% 2-propanol
250
-
5-Cholesten-3-one
-
isomerization to 4-cholesten-3-one, rapid reaction, stopped-flow measurement, 500 mM phosphate buffer, pH 7.5, in the presence of 1% thesit and 1.25% 2-propanol
278
-
5-Cholesten-3-one
-
assay relies on spectroscopic detection of 4-cholesten-3-one formation
330
-
5-Cholesten-3-one
-
isomerization to 4-cholesten-3-one, rapid reaction, stopped-flow measurement, 500 mM phosphate buffer, pH 7.5, in the presence of 1% thesit and 1.25% 2-propanol
332
-
5-Cholesten-3-one
-
assay relies on spectroscopic detection of 4-cholesten-3-one formation
670
-
5-Cholesten-3-one
-
isomerization to 4-cholesten-3-one, rapid reaction, stopped-flow measurement, 50 mM phosphate buffer, pH 7.5, in the presence of 1% thesit and 1.25% 2-propanol
37
-
cholestanol
-
assay relies on detection of H2O2 formation
37
-
cholestanol
-
H2O2 production assayed, 100 mM phosphate buffer
37
-
cholestanol
-
substrate cholestanol, rate of H2O2 formation detected with o-dianisidine and horseradish peroxidase. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
40
-
cholestanol
-
assay relies on detection of H2O2 formation
40
-
cholestanol
-
H2O2 production assayed, 100 mM phosphate buffer
40
-
cholestanol
-
substrate cholestanol, rate of H2O2 formation detected with o-dianisidine and horseradish peroxidase. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
0.0014
-
cholesterol
-
mutant enzyme H447E/E361Q
0.017
-
cholesterol
-
mutant enzyme H447E
0.035
-
cholesterol
-
mutant enzyme N485D, determined by H2O2 detection
0.035
-
cholesterol
-
apparent value, mutant enzyme N485L, pH 5.1, temperature not specified in the publication
0.044
-
cholesterol
-
apparent value, mutant enzyme N485L, pH 7.0, temperature not specified in the publication
0.073
-
cholesterol
-
mutant enzyme N485D, determined by cholest-4-en-3-one detection
0.073
-
cholesterol
-
apparent value, mutant enzyme N485D, pH 7.0, temperature not specified in the publication
0.093
-
cholesterol
-
mutant enzyme H447Q/E361Q
0.32
-
cholesterol
-
mutant enzyme H447Q
0.55
-
cholesterol
-
H69A mutant enzyme, steady state, 50 mM phosphate buffer, 1% thesit, 1% 2-propanol
0.8
-
cholesterol
-
assay relies on spectroscopic detection of 4-cholesten-3-one formation, decreasing length of C17 chain affects turnover negatively
0.85
-
cholesterol
-
mutant enzyme G347N, determined by H2O2 detection
0.86
-
cholesterol
-
mutant enzyme F359W, determined by cholest-4-en-3-one detection
1
-
cholesterol
-
mutant enzyme G347N, determined by cholest-4-en-3-one detection
1.3
-
cholesterol
-
mutant enzyme F359W, determined by H2O2 detection
1.4
-
cholesterol
-
mutant enzyme E361
1.4
-
cholesterol
-
apparent value, mutant enzyme N485D, pH 5.1, temperature not specified in the publication
1.6
-
cholesterol
-
H69A mutant enzyme, cholesterol oxidation, 50 mM phosphate buffer, 1% thesit, 1% 2-propanol
2.21
-
cholesterol
-
recombinant mutant V191A, pH 7.0, 22C
3
-
cholesterol
-
substrate cholesterol, polarographic determination of the rate of oxygen consumption. 1% thesit (polyoxyethylene(9)-lauryl-ether), 10% propan-2-ol and 50 mM phosphate, 25C and pH 7.5
6
-
cholesterol
-
substrate cholesterol, polarographic determination of the rate of oxygen consumption. 1% thesit (polyoxyethylene(9)-lauryl-ether), 10% propan-2-ol and 50 mM phosphate, 25C and pH 7.5
9
-
cholesterol
-
pH 7.0, 37C, mutant enzyme P357N
9
-
cholesterol
-
substrate cholesterol, polarographic determination of the rate of oxygen consumption. 0.5 M potassium phosphate, 1% thesit (polyoxyethylene(9)-lauryl-ether), 1.25% propan-2-ol, 25C and pH 7.5
11
-
cholesterol
-
assay relies on spectroscopic detection of 4-cholesten-3-one formation
11
-
cholesterol
-
pH 7.0, 37C, mutant enzyme V145D; pH 7.0, 37C, mutant enzyme V145Q
11
-
cholesterol
-
substrate cholesterol, detection of product formation (cholest-4-en-3-one) at 240 nm. 0.5 M potassium phosphate, 1% thesit (polyoxyethylene(9)-lauryl-ether), 1.25% propan-2-ol, 25C and pH 7.5
15
-
cholesterol
-
pH 7.0, 37C, mutant enzyme L119A
23
-
cholesterol
-
pH 7.0, 37C, mutant enzyme Q286R; pH 7.0, 37C, mutant enzyme V145E
27
-
cholesterol
-
pH 7.0, 37C, mutant enzyme L119F
28
-
cholesterol
-
50 mM phosphate buffer, pH 7.5
29
-
cholesterol
-
pH 7.0, 37C, mutant enzyme S379T
32
-
cholesterol
-
substrate cholesterol, rate of H2O2 formation detected with o-dianisidine and horseradish peroxidase. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
40
-
cholesterol
-
pH 7.0, 37C, wild-type enzyme
42
-
cholesterol
-
wild-type enzyme, determined by cholest-4-en-3-one detection
43
-
cholesterol
-
substrate cholesterol, polarographic determination of the rate of oxygen consumption. 1% thesit (polyoxyethylene(9)-lauryl-ether), 10% propan-2-ol and 50 mM phosphate, 25C and pH 7.5
44
-
cholesterol
-
-
44
-
cholesterol
-
wild-type enzyme
46
-
cholesterol
-
apparent value, wild type enzyme, pH 5.1, temperature not specified in the publication
47
-
cholesterol
-
wild-type enzyme, determined by H2O2 detection
47
-
cholesterol
-
apparent value, wild type enzyme, pH 7.0, temperature not specified in the publication
47.3
-
cholesterol
-
recombinant wild-type enzyme, pH 7.0, 22C
48
-
cholesterol
-
50 mM phosphate buffer, pH 7.5
48
-
cholesterol
-
substrate cholesterol, detection of product formation (cholest-4-en-3-one) at 240 nm. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5; substrate cholesterol, rate of H2O2 formation detected with o-dianisidine and horseradish peroxidase. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
48.1
-
cholesterol
-
product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer
49
-
cholesterol
-
pH 7.0, 37C, mutant enzyme S379A
51
-
cholesterol
-
pH 7.0, 37C, mutant enzyme S379V
56
-
cholesterol
-
substrate cholesterol, polarographic determination of the rate of oxygen consumption. 0.5 M potassium phosphate, 1% thesit (polyoxyethylene(9)-lauryl-ether), 1.25% propan-2-ol, 25C and pH 7.5
57
-
cholesterol
-
substrate cholesterol, polarographic determination of the rate of oxygen consumption. 1% thesit (polyoxyethylene(9)-lauryl-ether), 10% propan-2-ol and 50 mM phosphate, 25C and pH 7.5
63
-
cholesterol
-
product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer
63
-
cholesterol
-
substrate cholesterol, detection of product formation (cholest-4-en-3-one) at 240 nm. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
67
-
cholesterol
-
assay relies on spectroscopic detection of 4-cholesten-3-one formation
67
-
cholesterol
-
substrate cholesterol, detection of product formation (cholest-4-en-3-one) at 240 nm. 0.5 M potassium phosphate, 1% thesit (polyoxyethylene(9)-lauryl-ether), 1.25% propan-2-ol, 25C and pH 7.5
105
-
cholesterol
-
50 mM phosphate buffer, pH 7.5, 1% thesit, 10% 2-propanol
345
-
cholesterol
-
50 mM phosphate buffer, pH 7.5, 1% thesit, 10% 2-propanol
0.69
-
dehydroepiandrosterone
-
-
7
-
epiandrosterone
-
H2O2 production assayed, 100 mM phosphate buffer
9
-
pregnenolone
-
pH 7.0, 37C, mutant enzyme P357N; pH 7.0, 37C, mutant enzyme V145D
11
-
pregnenolone
-
pH 7.0, 37C, mutant enzyme V145E
12
-
pregnenolone
-
pH 7.0, 37C, mutant enzyme V145Q
13
-
pregnenolone
-
pH 7.0, 37C, mutant enzyme L119A
19
-
pregnenolone
-
pH 7.0, 37C, mutant enzyme Q286R
21
-
pregnenolone
-
assay relies on spectroscopic detection of 4-cholesten-3-one formation
21
-
pregnenolone
-
product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer
21
-
pregnenolone
-
pH 7.0, 37C, mutant enzyme L119F
21
-
pregnenolone
-
substrate pregnenolone, detection of product formation at 240 nm. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
21
-
pregnenolone
-
substrate pregnenolone, rate of H2O2 formation detected with o-dianisidine and horseradish peroxidase. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
24
-
pregnenolone
-
assay relies on spectroscopic detection of 4-cholesten-3-one formation
24
-
pregnenolone
-
product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer
24
-
pregnenolone
-
substrate pregnenolone, detection of product formation at 240 nm. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
30
-
pregnenolone
-
pH 7.0, 37C, wild-type enzyme
35
-
pregnenolone
-
pH 7.0, 37C, mutant enzyme S379A
35
-
pregnenolone
-
substrate pregnenolone, rate of H2O2 formation detected with o-dianisidine and horseradish peroxidase. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
39
-
pregnenolone
-
pH 7.0, 37C, mutant enzyme S379V
45
-
pregnenolone
-
pH 7.0, 37C, mutant enzyme S379T
0.8
-
trans-androsterone
-
substrate trans-androsterone, rate of H2O2 formation detected with o-dianisidine and horseradish peroxidase. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
7
-
trans-androsterone
-
substrate trans-androsterone, rate of H2O2 formation detected with o-dianisidine and horseradish peroxidase. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
0.8
-
trans-dehydroandrosterone
-
substrate trans-dehydroandrosterone, detection of product formation at 240 nm. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
1
-
trans-dehydroandrosterone
-
substrate trans-dehydroandrosterone, rate of H2O2 formation detected with o-dianisidine and horseradish peroxidase. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
6
-
trans-dehydroandrosterone
-
substrate trans-dehydroandrosterone, rate of H2O2 formation detected with o-dianisidine and horseradish peroxidase. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
8.2
-
trans-dehydroandrosterone
-
substrate trans-dehydroandrosterone, detection of product formation at 240 nm. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25C and pH 7.5
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0013
-
cholesterol
-
apparent value, silk mat-immobilized enzyme, in 50 mM potassium phosphate buffer, pH 7.5, 30C
8550
394
-
cholesterol
-
recombinant wild-type enzyme, pH 7.0, 22C
8550
902
-
cholesterol
-
recombinant mutant V191A, pH 7.0, 22C
8550
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.381
-
O2
-
mutant enzyme F359W
1.383
-
O2
-
wild-type enzyme
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.9
-
-
isoelectric focusing with carrier ampholites pH 4-6, cholesterol oxidase II
5.7
-
C5II17, -
calculated from amino acid sequence
7
-
-
isoelectric fucusing, pH-gradient 3.5-10.0
8.5
-
A7LGL0
calculated from amino acid sequence
9
-
Q157H4
pI-value about 9.0, isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Nocardia erythropolis
-
-
Manually annotated by BRENDA team
-
the strain reaches its maximal utilization of cholesterol as the only C source for production of extracellular cholesterol oxidase
Manually annotated by BRENDA team
additional information
-
enzyme production reaches its maximum by incubation at 30C for 12 d
Manually annotated by BRENDA team
additional information
-
Streptomyces parvus shows markedly high cho activity in culture broth
Manually annotated by BRENDA team
additional information
-
effect of carbon and nitrogen sources on COD production, highest activity on lactose and sucrose and on yeast extract, respectively, overview
Manually annotated by BRENDA team
additional information
Streptomyces parvus 13647 J
-
Streptomyces parvus shows markedly high cho activity in culture broth
-
Manually annotated by BRENDA team
additional information
Streptomyces sp. R-6
-
effect of carbon and nitrogen sources on COD production, highest activity on lactose and sucrose and on yeast extract, respectively, overview
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
H9NJ43, H9NJ52
; ChoM1 is mainly associated with the cell envelope
Manually annotated by BRENDA team
-
produces both cell-linked and extracellular enzyme in high amount
-
Manually annotated by BRENDA team
-
produces both cell-linked and extracellular enzyme in high amount
-
Manually annotated by BRENDA team
Pseudomonas spp.
-
-
-
Manually annotated by BRENDA team
Streptomyces violascens, Streptoverticillum cholesterolicum
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Manually annotated by BRENDA team
H9NJ43, H9NJ52
; ChooM2 is mainly extracellular and soluble
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Manually annotated by BRENDA team
Bacillus sp. SFF34, Bordetella sp. B4, Rhodococcus sp. CECT3014, Streptomyces parvus 13647 J, Streptomyces sp. R-6
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Manually annotated by BRENDA team
Arthrobacter sp., Mycobacterium sp., Nocardia erythropolis, Nocardia rhodochrous, Rhodococcus equi
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Manually annotated by BRENDA team
Arthrobacter sp., Mycobacterium sp., Nocardia erythropolis, Nocardia rhodochrous, Rhodococcus equi
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Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
Streptomyces sp. (strain SA-COO)
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
Nocardia rhodochrous
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3 enzyme forms differ chiefly in the presence or absence of hydrophobic anchor region connected by a trypsin-sensitive region, no phospholipids are extracted with the enzyme
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystals of recombinant His-tagged enzyme, vapor diffusion by hanging drop, space group P21
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rod shaped crystals with bright yellow color
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structure of substrate-enzyme complex, 1.8 A resolution, substrate binds in internal cavity which is totally sealed from solvent
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hanging-drop vapour-diffusion method using polyethylene glycol as a precipitating agent. The crystals belong to space group P3(1)21 with unit-cell parameters a = b = 119.6, c = 101.1 A, and has one subunit in the asymmetric unit
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crystal screen based on sparse matrix formulation, using 500 mM ammonium sulfate, 1.0 M lithium sulfate and 10 mM sodium citrate
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crystals of the H447Q/E361Q mutant protein are grown by hanging-drop vapor diffusion method, high-resolution crystal structure of mutant enzyme H447Q/E361Q with glycerol and without glycerol
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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
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structure of the F359W mutant enzyme, hanging drop vapor diffusion method. In the atomic-resolution structure of F359W, the indole ring of the tryptophan completely fills the tunnel and is observed in only a single conformation
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Sub-atomic resolution crystal structure, crystallographic data: space group P2(1), a = 52.23 A, b * 72.9 A, c = 62.95 A, beta = 105.1
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vapor diffusion by hanging drop method, 1.5 A resolution, wild-type, E361Q, H447N and H447Q mutant, structure consists of FAD-binding and a steroid-binding domain with a large active site cavity in between
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ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Acetone
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37C, 24 h, 1 ml organic solvent added to 2 ml culture filtrate, 90% loss of activity
Acetone
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the enzyme is unstable in 50% (v/v) acetone; the enzyme remains stable after incubation in 50% (v/v) for 24 h at 37C
Acetone
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6% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
Acetone
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6% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
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benzene
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37C, 24 h, 1 ml organic solvent added to 2 ml culture filtrate, stable
benzene
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127% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
Butanol
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37C, 24 h, 1 ml organic solvent added to 2 ml culture filtrate, stable
Butanol
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the enzyme remains stable after incubation in 50% (v/v) for 24 h at 37C
Butanol
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77% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
Butanol
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77% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
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chloroform
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37C, 24 h, 1 ml organic solvent added to 2 ml culture filtrate, stable
chloroform
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96% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
cyclohexane
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119% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
cyclooctane
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37C, 24 h, 1 ml organic solvent added to 2 ml culture filtrate, stable
dimethyl sulfoxide
-
37C, 24 h, 1 ml organic solvent added to 2 ml culture filtrate, stable
dimethyl sulfoxide
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88% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
DMSO
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the enzyme remains stable after incubation in 50% (v/v) for 24 h at 37C
Emal 20CM
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the enzyme is relatively tolerant to treatment with Emal 20CM even at 60C
Ethanol
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37C, 24 h, 1 ml organic solvent added to 2 ml culture filtrate, stable
Ethanol
-
the enzyme remains stable after incubation in 50% (v/v) for 24 h at 37C
Ethanol
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5% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
Ethanol
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5% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
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Ethyl acetate
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37C, 24 h, 1 ml organic solvent added to 2 ml culture filtrate, stable
Ethyl acetate
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the enzyme remains stable after incubation in 50% (v/v) for 24 h at 37C
Ethyl acetate
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93% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
Ethyl acetate
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93% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
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isopropanol
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37C, 24 h, 1 ml organic solvent added to 2 ml culture filtrate, stable
isopropanol
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the enzyme remains stable after incubation in 50% (v/v) for 24 h at 37C
Methanol
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37C, 24 h, 1 ml organic solvent added to 2 ml culture filtrate, stable
Methanol
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the enzyme remains stable after incubation in 50% (v/v) for 24 h at 37C
Methanol
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4% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
p-xylene
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37C, 24 h, 1 ml organic solvent added to 2 ml culture filtrate, stable
propan-2-ol
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the activity of the enzyme is rapidly inactivated in the presence of 30% (v/v) propan-2-ol at 25C
SDS
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the enzyme is relatively tolerant to treatment with SDS after incubation for 1 h at 30C
sodium cholate
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the enzyme is stable at 50% (v/v) sodium cholate after incubation for 1 h at 30C
sodium cholate
A9QAE7
the enzyme is stable in sodium cholate after incubation for 1 h at 30C
sodium cholate
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the enzyme retains more than 80% of its original activity in 0.5% (v/v) sodium cholate after incubation for 1 h at 60C
sodium cholate
uncultured gamma proteobacterium Y-134
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the enzyme retains more than 80% of its original activity in 0.5% (v/v) sodium cholate after incubation for 1 h at 60C
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sodium dodecyl sarcosinate
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the enzyme is relatively tolerant to treatment with sodium dodecyl sarcosinate even at 60C
sodium laurylbenzenesulfonate
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the enzyme is relatively tolerant to treatment with sodium laurylbenzenesulfonate after incubation for 1 h at 30C
sodium polyoxyethylene alkyl ether sulfate
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the enzyme is stable at 50% (v/v) sodium polyoxyethylene alkyl ether sulfate after incubation for 1 h at 30C
sodium polyoxyethylene alkyl ether sulfate
A9QAE7
the enzyme is stable in sodium polyoxyethylene alkyl ether sulfate after incubation for 1 h at 30C
toluene
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37C, 24 h, 1 ml organic solvent added to 2 ml culture filtrate, stable
Triton X-100
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the enzyme is stable at 50% (v/v) Triton X-100 after incubation for 1 h at 30C
Triton X-100
A9QAE7
the enzyme is stable in Triton X-100 after incubation for 1 h at 30C
Triton X-405
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the enzyme is stable at 50% (v/v) Triton X-405 after incubation for 1 h at 30C
Triton X-405
A9QAE7
the enzyme is stable in Triton X-405 after incubation for 1 h at 30C
Triton X-405
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the enzyme retains more than 80% of its original activity in 0.5% (v/v) Triton X-405 after incubation for 1 h at 60C
Triton X-405
uncultured gamma proteobacterium Y-134
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the enzyme retains more than 80% of its original activity in 0.5% (v/v) Triton X-405 after incubation for 1 h at 60C
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Tween 20
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the enzyme is stable at 50% (v/v) Tween 20 after incubation for 1 h at 30C
Tween 20
A9QAE7
the enzyme is stable in Tween 20 after incubation for 1 h at 30C
urea
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the apoprotein of the mutant H69A lacks the characteristic tertiary structure of the holoprotein and displays larger hydrophobic surfaces. Its urea-induced unfolding does not occur by a simple two-state mechanism and is largely nonreversible. Minor alterations in the flavin binding region are evident between the native and the refolded proteins, and are likely responsible for the low refolding yield observed
xylene
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93% the activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
Methanol
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4% activity remaining, pH 7.0, 37C, 24 h, with mechanical shaking
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additional information
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the enzyme shows moderate stability towards all organic solvents except acetone, benzene and chloroform. The activity increases in presence of isopropanol and ethanol
additional information
Streptomyces sp. R-6
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the enzyme shows moderate stability towards all organic solvents except acetone, benzene and chloroform. The activity increases in presence of isopropanol and ethanol
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ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
E311D
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mutations of Glu311 cause a switch in the basic kinetic mechanism of the reaction of reduced cholesterol oxidase with dioxygen: wild-type cholesterol oxidase shows a saturation behavior with increasing oxygen concentration, while for Glu311 mutants a linear dependence is found that is assumed to reflect a simple second order process
E311L
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mutations of Glu311 cause a switch in the basic kinetic mechanism of the reaction of reduced cholesterol oxidase with dioxygen: wild-type cholesterol oxidase shows a saturation behavior with increasing oxygen concentration, while for Glu311 mutants a linear dependence is found that is assumed to reflect a simple second order process
E311Q
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mutations of Glu311 cause a switch in the basic kinetic mechanism of the reaction of reduced cholesterol oxidase with dioxygen: wild-type cholesterol oxidase shows a saturation behavior with increasing oxygen concentration, while for Glu311 mutants a linear dependence is found that is assumed to reflect a simple second order process
H69A
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exchange prevents formation of histidyl-FAD bond
H69A
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site-directed mutagenesis, the mutant does not bind FAD, the mutant is more sensitive to urea and unfolds at lower urea concentrations of 3 M compared to the wild-type enzyme at 5 M, the mutant also has a lower melting temperature
E361A
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site-directed mutagenesis, the mutant shows no dehydrogenase activity
E361Q
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H447Q mutant shows wild-type activity in isomerization, kcat for oxidation reaction is reduced 120fold, Km increases 2fold compared to wild-type, E361Q mutant has no isomerization activity, kcat is 30fold slower than for wild-type, Glu361 may therefore act as general base in oxidation reaction
E361Q
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turnover number for cholesterol is decreased 31.4fold compared to wild-type enzyme, 1.8fold increase in Km-value for cholesterol compared to wild-type enzyme
F359A
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site-directed mutagenesis, the dehydrogenase/oxidase ratio is 12fold increased compared with the ratio for the wild-type enzyme
F359W
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kcat for the F359W mutant-catalyzed reaction decreases 13fold relative to that of the wild-type-catalyzed reaction. Transfer of hydride from the sterol to the flavin prosthetic group is no longer rate-limiting for the mutant enzyme. Kinetic cooperativity with respect to molecular oxygen is observed with the tunnel mutant, but not with the wild-type enzyme. In the atomic-resolution structure of F359W, the indole ring of the tryptophan completely fills the tunnel and is observed in only a single conformation. The size of the indole is proposed to limit conformational rearrangement of residue 359 that leads to tunnel opening in the wild-type enzyme
F444A
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site-directed mutagenesis, the dehydrogenase/oxidase ratio is 4fold increased compared with the ratio for the wild-type enzyme
G347N
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mutant can not be saturated with oxygen. Transfer of hydride from the sterol to the flavin prosthetic group is no longer rate-limiting for the mutant enzyme. Kinetic cooperativity with respect to molecular oxygen is observed with the tunnel mutant, but not with the wild-type enzyme
H447E
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turnover number for cholesterol is decreased about 2600fold compared to wild-type enzyme
H447E/E361Q
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turnover number for cholesterol is decreased about 31500fold compared to wild-type enzyme. Mutant enzyme is not able to isomerize cholest-5-en-3-one. Mutant enzyme is folded like native enzyme and still associates with model membranes
H447N
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H447Q mutant shows wild-type activity in isomerization, kcat for oxidation reaction is reduced 120fold, Km increases 2fold compared to wild-type, E361Q mutant has no isomerization activity, kcat is 30fold slower than for wild-type, Glu361 may therefore act as general base in oxidation reaction
H447Q
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H447Q mutant shows wild-type activity in isomerization, kcat for oxidation reaction is reduced 120fold, Km increases 2fold compared to wild-type, E361Q mutant has no isomerization activity, kcat is 30fold slower than for wild-type, Glu361 may therefore act as general base in oxidation reaction
H447Q
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turnover number for cholesterol is decreased 137.5fold compared to wild-type enzyme, Km-value for cholesterol is identical to wild-type value
H447Q/E361Q
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turnover number for cholesterol is decreased 473fold compared to wild-type enzyme, 1.7fold increase in Km-value for cholesterol compared to wild-type enzyme. Mutant enzyme is not able to isomerize cholest-5-en-3-one
L117P
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mutant enzyme without activity
L119A
-
specific activity is similar to that of the wild-type enzyme
L119F
-
specific activity is similar to that of the wild-type enzyme
N485A
-
site-directed mutagenesis, the mutant shows no dehydrogenase activity
N485D
-
mutant can not be saturated with oxygen. Transfer of hydride from the sterol to the flavin prosthetic group is no longer rate-limiting for the mutant enzyme. Kinetic cooperativity with respect to molecular oxygen is observed with the tunnel mutant, but not with the wild-type enzyme
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
N485L
-
the kcat of N485D is diminished about 1110times compared with that of wild type, while the apparent Km value is minimally affected
P357N
-
specific activity is similar to that of the wild-type enzyme
Q286R
-
specific activity is similar to that of the wild-type enzyme
R135H
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random mutagenesis
S103T
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random mutagenesis
S279R
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mutant enzyme without activity on cholesterol and pregnenolone
S379D
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mutant enzyme without activity on cholesterol and pregnenolone
S379T
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specific activity is similar to that of the wild-type enzyme
V121A
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random mutagenesis
V124A
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site-directed mutagenesis, the dehydrogenase/oxidase ratio is 30fold increased compared with the ratio for the wild-type enzyme
V145E
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random mutagenesis
V145Q
-
specific activity is similar to that of the wild-type enzyme
V191A
-
site-directed mutagenesis, the mutant enzymes shows a significant decrease in its oxidase activity, but shows increased dehydrogenase activity. The dehydrogenase/oxidase ratio of Val191Ala is more than 150%, which is a 408fold increase compared with the ratio for the wild-type enzyme, substrate inhibition with cholesterol
Y446A
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site-directed mutagenesis, the dehydrogenase/oxidase ratio is 3fold increased compared with the ratio for the wild-type enzyme
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
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N485L
-
the kcat of N485D is diminished about 1110times compared with that of wild type, while the apparent Km value is minimally affected
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R135H
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random mutagenesis
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S103T
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random mutagenesis
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V121A
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random mutagenesis
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H69A
-
mutation results in a significant decrease in activity, in the midpoint redox potential of the flavin, and in stability with respect to the wild-type enzyme, but does not modify the overall structure of the enzyme
additional information
-
development and evaluation of a cholesterol biosensor based on immobilized cholesterol esterase and cholesterol oxidase on oxygen electrode for the determination of total cholesterol in food samples
additional information
H9NJ43, H9NJ52
development of a strategy to overcome the rate-limiting step catalzed by the enzyme by augmenting the activity of cholesterol oxidases in Mycobacterium neoaurum strains to enhance their transformation productivity of sterols to valuable steroids, overview; development of a strategy to overcome the rate-limiting step catalzed by the enzyme by augmenting the activity of cholesterol oxidases in Mycobacterium neoaurum strains to enhance their transformation productivity of sterols to valuable steroids, overview
additional information
-
engineered ChOx to improve activity and alter specifity
M122A
-
site-directed mutagenesis, the dehydrogenase/oxidase ratio is 26fold increased compared with the ratio for the wild-type enzyme
additional information
-
site-directed mutagenesis on oxygen-binding residues, which are observed in the high-resolution crystal structure, in order to elucidate the amino acid residues responsible for the oxidase activity, overview. Engineering of the enzyme for electrochemical monitoring of cholesterol
V145E
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random mutagenesis
-
additional information
Streptomyces violascens, Streptoverticillum cholesterolicum
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engineered ChOx to improve activity and alter specificity
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
unfolding and folding after urea treatment of wild-type enzyme and mutant H69A, equilibrium unfolding study, kinetics
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