Information on EC 1.14.13.141 - cholest-4-en-3-one 26-monooxygenase [(25S)-3-oxocholest-4-en-26-oate forming]

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.13.141
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RECOMMENDED NAME
GeneOntology No.
cholest-4-en-3-one 26-monooxygenase [(25S)-3-oxocholest-4-en-26-oate forming]
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(25S)-26-hydroxycholest-4-en-3-one + NADH + H+ + O2 = (25S)-26-oxocholest-4-en-3-one + NAD+ + 2 H2O
show the reaction diagram
(1b)
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(25S)-26-oxocholest-4-en-3-one + NADH + H+ + O2 = (25S)-3-oxocholest-4-en-26-oate + NAD+ + H2O
show the reaction diagram
(1c)
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-
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cholest-4-en-3-one + 3 NADH + 3 H+ + 3 O2 = (25S)-3-oxocholest-4-en-26-oate + 3 NAD+ + 4 H2O
show the reaction diagram
overall reaction
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-
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cholest-4-en-3-one + NADH + H+ + O2 = (25S)-26-hydroxycholest-4-en-3-one + NAD+ + H2O
show the reaction diagram
(1a)
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Steroid degradation
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SYSTEMATIC NAME
IUBMB Comments
cholest-4-en-3-one,NADH:oxygen oxidoreductase [(25S)-3-oxocholest-4-en-26-oate forming]
This enzyme, found in several bacterial pathogens, is involved in degradation of the host's cholesterol. It catalyses the hydroxylation of the C-26 carbon, followed by oxidation of the alcohol to the carboxylic acid via the aldehyde intermediate, initiating the degradation of the alkyl side-chain of cholesterol [4]. The products are exclusively in the (25S) configuration. It is a two-component system consisting of a P-450 (heme thiolate) oxygenase (Cyp125) and a ferredoxin reductase (most likely KshB, which is also a part of EC 1.14.13.142, 3-ketosteroid 9alpha-monooxygenase). The enzyme also accepts cholesterol as a substrate. cf. EC 1.14.13.221, cholest-4-en-3-one 27-monooxygenase.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
27-hydroxycholest-4-en-3-one + O2
cholest-4-en-3-one-27-oic acid + H2O
show the reaction diagram
-
-
-
?
cholest-4-en-3-one + O2 + NADH + H+
(25S)-26-hydroxycholest-4-en-3-one + H2O + NAD+
show the reaction diagram
cholest-4-en-3-one + O2 + NADH + H+
26-hydroxycholest-4-en-3-one + H2O + NAD+
show the reaction diagram
cholest-4-en-3-one + O2 + NADPH + H+
27-hydroxycholest-4-en-3-one + H2O + NADP+
show the reaction diagram
physiological substrate, the enzyme hydroxylates cholest-4-en-3-one at C-27
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-
?
cholesterol + O2 + NADH + H+
?
show the reaction diagram
cholesterol + O2 + NADPH + H+
27-hydroxycholesterol + H2O + NADP+
show the reaction diagram
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-
-
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
27-hydroxycholest-4-en-3-one + O2
cholest-4-en-3-one-27-oic acid + H2O
show the reaction diagram
P9WPP0
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-
-
?
cholest-4-en-3-one + O2 + NADPH + H+
27-hydroxycholest-4-en-3-one + H2O + NADP+
show the reaction diagram
P9WPP0
physiological substrate, the enzyme hydroxylates cholest-4-en-3-one at C-27
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-
?
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
alpha-[(4-methylcyclohexyl)carbonyl amino]-N-4-pyridinyl-1H-indole-3-propanamide
LP10, 30% inhibition at 0.2 mM
nitric oxide
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reversible inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0208
cholest-4-en-3-one
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pH and temperature not specified in the publication
0.0107
cholesterol
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pH and temperature not specified in the publication
1.2
O2
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apparent value, Km above 1.2 mM, in 0.1 M potassium phosphate at pH 7.0, temperature not specified in the publication
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.92
cholest-4-en-3-one
Mycobacterium tuberculosis
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pH and temperature not specified in the publication
0.47
cholesterol
Mycobacterium tuberculosis
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pH and temperature not specified in the publication
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
11000
O2
Mycobacterium tuberculosis
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apparent value, in 0.1 M potassium phosphate at pH 7.0, temperature not specified in the publication
9
PDB
SCOP
CATH
ORGANISM
UNIPROT
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
46400
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MALDI-TOF mass spectrometry
48200
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MALDI-TOF mass spectrometry
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 47200, calculated from amino acid sequence
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
enzyme bound to cholest-4-en-3-one, hanging drop vapor diffusion method, using 0.2 M ammonium sulfate, 0.1 M bis-Tris, pH 5.5, and 25% (w/v) PEG 3350
hanging drop vapor diffusion method, using 2.0 M ammonium sulfate, 0.1 M bis-Tris (pH 5.5) and 0.5 M NaCl (truncated enzyme C429L in complex with inhibitor) or 0.1 M ammonium acetate, 0.1 M bis-Tris (pH 5.5) and 17% PEG 10000, or 2.0 M ammonium sulfate and 0.1 M Tris HCl pH 8.5 (truncated enzyme C429L without any ligand)
sitting drop vapor diffusion method, using MgCl2 with 0.1 M HEPES, pH 7.0 or 7.5, and 20% (w/v) PEG 6000 or 25% (w/v) PEG 3350, respectively
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ni-NTA resin column chromatography, Resource-Q column chromatography, and Sephacryl S-200 gel filtration
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Ni2+-affinity column chromatography and Q Sepharose column chromatography
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Ni2+-NTA column chromatography
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Source 15Q column chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
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expressed in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli DH5alpha cells
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expressed in Escherichia coli HMS174 (DE3) cells
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expressed in Rhodococcus jostii strain RHA1
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
cyp125 is up-regulated 7.1fold with growth on cholesterol
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE