Information on EC 1.13.11.48 - 3-hydroxy-2-methylquinolin-4-one 2,4-dioxygenase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.13.11.48
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RECOMMENDED NAME
GeneOntology No.
3-hydroxy-2-methylquinolin-4-one 2,4-dioxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
3-hydroxy-2-methyl-1H-quinolin-4-one + O2 = N-acetylanthranilate + CO
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
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redox reaction
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-
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reduction
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SYSTEMATIC NAME
IUBMB Comments
3-hydroxy-2-methyl-1H-quinolin-4-one 2,4-dioxygenase (CO-forming)
Does not contain a metal centre or organic cofactor. Fission of two C-C bonds: 2,4-dioxygenolytic cleavage with concomitant release of carbon monoxide. The enzyme from Arthrobacter sp. can also act on 3-hydroxy-4-oxoquinoline, forming N-formylanthranilate and CO (cf. EC 1.13.11.47, 3-hydroxy-4-oxoquinoline 2,4-dioxygenase), but more slowly.
CAS REGISTRY NUMBER
COMMENTARY hide
160995-63-1
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1H-3-hydroxy-4-oxoquinaldine + O2
?
show the reaction diagram
1H-3-Hydroxy-4-oxoquinaldine + O2
N-Acetylanthranilate + CO
show the reaction diagram
1H-3-hydroxy-4-oxoquinaldine + O2
N-acetylanthranilic acid + CO
show the reaction diagram
1H-3-hydroxy-4-oxoquinoline + O2
N-formylanthranilic acid + CO
show the reaction diagram
3-hydroxy-1H-quinoline-4-one + O2
N-formylanthranilate + CO
show the reaction diagram
3-hydroxy-2-methyl-1H-quinolin-4-one + O2
N-acetylanthranilate + CO
show the reaction diagram
additional information
?
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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
1H-3-hydroxy-4-oxoquinaldine + O2
?
show the reaction diagram
1H-3-Hydroxy-4-oxoquinaldine + O2
N-Acetylanthranilate + CO
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
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no cofactors
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ca2+
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1 mM, weak inhibition
Co2+
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1 mM, weak inhibition
Cu2+
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1 mM, strong inhibition
Fe2+
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1 mM, strong inhibition
guanidine hydrochloride
almost linear decrease of activity up to 1 M where activity vanishes, three-state unfolding of mutant enzyme; causes isothermal unfolding of mutant C69S in a three-state mechanism, thermodynamic analysis, overview
Mg2+
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1 mM, weak inhibition
Mn2+
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1 mM, weak inhibition
Ni2+
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1 mM, strong inhibition
Urea
causes isothermal unfolding of mutant C69S in a three-state mechanism, thermodynamic analysis, overview; nonlinear decrease of activity, activity lost at about 5 M, three-state unfolding of mutant enzyme
Zn2+
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1 mM, strong inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0092 - 0.1907
1H-3-Hydroxy-4-oxoquinaldine
0.0017
3-hydroxy-2-methyl-1H-quinolin-4-one
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30°C, pH 8.0
0.325 - 1.646
O2
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.7 - 28.2
1H-3-Hydroxy-4-oxoquinaldine
35 - 145
3-hydroxy-2-methyl-1H-quinolin-4-one
0.037 - 113.7
O2
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
70
sodium phosphate buffer, 30°C, pH 7.5
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.1 - 9.1
pH profile, C59S Hod, overview
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
N-terminally His6-tagged HOD is crystallized by the hanging-drop vapour-diffusion method using sodium/potassium tartrate as a precipitant and CuCl2 as an additive. The structure is solved by the single anomalous dispersion technique using data collected to 3.5 A resolution at the Cu absorption peak wavelength. The crystals belong to the primitive tetragonal space group P43212, with unit-cell parameters a = b = 153.788, c = 120.872 A; purified recombinant wild-type and mutants C69S and C69S/H251A N-terminally His6-tagged HOD, hanging drop vapour diffusion method, 50 mg/ml protein in 20 mM Tris-HCl pH 7.5, 100 mM NaCl, 2 mM EDTA, 1 mM DTT, is mixed with 1.65 M sodium/potassium tartrate, 0.1 M HEPES, pH 7.0, and 30 mM CuCl2, method optimization, X-ray diffraction structure determination and analysis at 3.5 A resolution, single anomalous dispersion technique
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 11
incubation for 5 min at pH between 5.5 and 11.0 does not result in significant loss of activity
673207
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10 - 40
enzyme in native reduced state (40 mM dithiothreitol) of mutant enzyme, 10 mM sodium phosphate and 10 mM sodium borate buffer, pH 7.5, unfolding starts at 40°C
10 - 50
native oxidized state of mutant enzyme, 10 mM sodium phosphate and 10 mM sodium borate buffer, pH 7.5, unfolding starts at 50°C
additional information
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recombinant His6HodC exhibits three-state unfolding with an intermediate state I that exhibits at the transition temperature a volume larger than that of the native or denatured state. The intermediate state I is also associated with the highest isothermal expansion coefficient, alphaP, of the three states and exhibits a significantly lower percentage of R-helical structure than the native state. The stability difference between the native and intermediate state is rather small which makes I a potential candidate for reactions with various ligands, particularly those having a preference for the apparently preserved beta-type motifs
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; recombinant mutant N-terminally His6-tagged HODs from Escherichia coli strain M15 by nickel affinity and anion exchange chromatography
recombinant wild-type and mutant N-terminally His6-tagged HODs from Escherichia coli strain M15 by nickel affinity and anion exchange chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli; sequence comparisons, overexpression of mutant N-terminally His6-tagged HODs in Escherichia coli strain M15
expression in Escherichia coli
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expression of wild-type and mutant N-terminally His6-tagged HODs in Escherichia coli strain M15
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overexpression of wild-type and mutant N-terminally His6-tagged HODs in Escherichia coli strain M15
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C69S/H251A
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site-directed mutagenesis, the mutant is catalytically inactive owing to the Ala substitution of the essential residue His251
E224A/C69S
site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme
F219Y
does not affect the kinetic parameters of the enzyme
F219Y/C69S
site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme
H102Q
Km for the heteroaromatic substrate is increased only 2.8fold, and kcat is reduced 2.3fold
H102Q/C69S
site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme
S101A
about 21fold increase in the Km for the organic substrate
S101A/C69S
site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme
S220N
does not drastically influence the kinetic parameters of the enzyme for the organic substrate, but it causes a 3.5fold decrease in Km for O2 and a 6.2fold decrease in kcat for O2
S220N/C69S
site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme
Y196A/C69S
site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme
Y196K/C69S
site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme
Y196R/C69S
site-directed mutagenesis, mutant apparent kinetic parameters compared to the wild-type enzyme
E224A
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decreased Km for O2
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H102Q
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Km for the heteroaromatic substrate is increased only 2.8fold, and kcat is reduced 2.3fold
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S101A
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about 21fold increase in the Km for the organic substrate
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C69S
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site-directed mutagenesis, the mutant has catalytic properties that are identical to those of wild-type HOD
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C69S/H251A
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site-directed mutagenesis, the mutant is catalytically inactive owing to the Ala substitution of the essential residue His251
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C69S/H251A
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site-directed mutagenesis, the mutant is catalytically inactive owing to the Ala substitution of the essential residue His251
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E224A
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decreased Km for O2
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H102Q
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Km for the heteroaromatic substrate is increased only 2.8fold, and kcat is reduced 2.3fold
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S101A
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about 21fold increase in the Km for the organic substrate
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