Information on EC 1.13.11.24 - quercetin 2,3-dioxygenase

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

EC NUMBER
COMMENTARY
1.13.11.24
-
RECOMMENDED NAME
GeneOntology No.
quercetin 2,3-dioxygenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
quercetin + O2 = 2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
-
-
-
quercetin + O2 = 2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
structure-function analysis of the active site
P42106
quercetin + O2 = 2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
EPR study, mechanism, N of His112 is the axial ligand of type II copper site
-
quercetin + O2 = 2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
hybrid density functional theory study on mechanism, dioxygen attack on copper is energetically preferred
-
quercetin + O2 = 2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
study on mobility and flexibility of substrate cavity, molecular dynamics simulations
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
rutin degradation
-
SYSTEMATIC NAME
IUBMB Comments
quercetin:oxygen 2,3-oxidoreductase (decyclizing)
The enzyme from Aspergillus sp. is a copper protein whereas that from Bacillus subtilis contains iron. Quercetin is a flavonol (5,7,3',4'-tetrahydroxyflavonol).
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
flavonol 2,4-dioxygenase
-
-
-
flavonol 2,4-oxygenase
-
-
-
-
manganese quercetin dioxygenase
-
-
pirin
-
-
QdoI
Bacillus subtilis 168
-
-
-
QueD
A2VA43
;
-
quercetin dioxygenase
-
-
quercetinase
-
-
-
-
quercetinase
-
-
quercetinase
Aspergillus flavus MTCC-1783, Aspergillus flavus MTCC-1883, Aspergillus flavus MTCC-1884, Aspergillus flavus MTCC-2206, Aspergillus flavus MTCC-2456
-
-
-
quercetinase
-
-
quercetinase
-
-
quercetinase
A7Y9J1
-
quercetinase
-
-
quercetinase
-
;
-
type III extradiol dioxygenase
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9075-67-6
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
PRL 1805
-
-
Manually annotated by BRENDA team
strain MTCC-1783; strain MTCC-1883; strain MTCC-1884; strain MTCC-2206; strain MTCC-2456
-
-
Manually annotated by BRENDA team
Aspergillus flavus MTCC-1783
strain MTCC-1783
-
-
Manually annotated by BRENDA team
Aspergillus flavus MTCC-1883
strain MTCC-1883
-
-
Manually annotated by BRENDA team
Aspergillus flavus MTCC-1884
strain MTCC-1884
-
-
Manually annotated by BRENDA team
Aspergillus flavus MTCC-2206
strain MTCC-2206
-
-
Manually annotated by BRENDA team
Aspergillus flavus MTCC-2456
strain MTCC-2456
-
-
Manually annotated by BRENDA team
Aspergillus flavus PRL 1805
PRL 1805
-
-
Manually annotated by BRENDA team
IFO-4408, grown on quercetin
-
-
Manually annotated by BRENDA team
DSM 821, grown on rutin
-
-
Manually annotated by BRENDA team
expression in Escherichia coli
-
-
Manually annotated by BRENDA team
Bacillus subtilis 168
-
-
-
Manually annotated by BRENDA team
medium improvement for production of enzyme
-
-
Manually annotated by BRENDA team
strain FLA
SwissProt
Manually annotated by BRENDA team
strain FLA, DSM 41951
-
-
Manually annotated by BRENDA team
strain FLA
SwissProt
Manually annotated by BRENDA team
strain FLA, DSM 41951
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
evolution
-
the ring-cleaving dioxygenase belongs to the cupin superfamily, characterized by a six-stranded beta-barrel fold and conserved amino acid motifs that provide the 3His or 2- or 3His-1Glu ligand environment of a divalent metal ion. The cupin domain comprises two conserved amino acid motifs with the consensus sequences G(X)5HXH(X)3-4E(X)6G (motif 1) and G(X)5-7PXG(X)2H(X)3N
evolution
-
the ring-cleaving dioxygenase belongs to the cupin superfamily, characterized by a six-stranded beta-barrel fold and conserved amino acid motifs that provide the 3His or 2- or 3His-1Glu ligand environment of a divalent metal ion. The cupin domain comprises two conserved amino acid motifs with the consensus sequences G(X)5HXH(X)3-4E(X)6G (motif 1) and G(X)5-7PXG(X)2H(X)3N
-
additional information
-
enzyme-bound quercetin shields the FeII cofactor from interactions with the O2 mimic nitric oxide, tentatively suggesting that the reaction catalyzed by Bacillus (Fe-)quercetinase may proceed without direct interaction of dioxygen and metal ion, overview
additional information
-
the mechanism which involves initial electron transfer from the divalent metal to O2, as proposed for the extradiol dioxygenases, requires that a MIII-superoxo state is thermodynamically accessible, overview
additional information
-
synthesis of a series of flavonolate complexes as structural and functional models for the enzyme-substrate complexes of the active site of MII-containing quercetin 2,3-dioxygenase, structure analysis, overview
additional information
-
Manual docking of the substrate quercetin into the active site showed that the different geometries of the copper site might be of catalytic importance
additional information
-
synthetic routes to tris(imidazolyl)carboxylate ligands, {N,N-bis[(1-methyl-1H-imidazol-4-yl-kappaN3)methyl]glycinato-kappa2N,O}(dichloro)ferrate(1-), [3-(hydroxy-kappaO)-2-phenyl-4H-chromen-4-onato-kappaO4][bis(triphenylphosphane)]copper, (1Z,3Z)-N,N'-di(pyridin-2-yl)-1H-isoindole-1,3(2H)-diimine, chloro[(4-phenoxy-1,4,7-triazonan-1-yl-kappa3N1,N4,N7)acetato-kappaO]copper,2-{4-[hydroxy(di-1H-imidazol-4-yl)methyl]-1H-imidazol-2-yl}-2-methylpropanoic acid, and 2-(4-{bis[1-methyl-2-(propan-2-yl)-1H-imidazol-4-yl]methyl}-1-methyl-1H-imidazol-2-yl)-2-methylpropanoic acid, and representative metal complexes, which provide more accurate active site models for several classes of redox metalloenzymes, complex structures, detailed overview
additional information
-
the mechanism which involves initial electron transfer from the divalent metal to O2, as proposed for the extradiol dioxygenases, requires that a MIII-superoxo state is thermodynamically accessible, overview
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3,5,7,2',4'-pentahydroxyflavone + O2
?
show the reaction diagram
-
5.5% of the activity with quercetin, Co-QueD, 0.9% of the activity with quercetin, Ni-QueD
-
-
?
3,5,7,3',4',5'-hexahydroxyflavone + O2
?
show the reaction diagram
-
77% of the activity with quercetin, Co-QueD, 46% of the activity with quercetin, Ni-QueD
-
-
?
3,5,7,4'-tetrahydroxyflavone + O2
?
show the reaction diagram
-
43% of the activity with quercetin, Co-QueD, 29% of the activity with quercetin, Ni-QueD
-
-
?
3,5,7-trihydroxyflavone + O2
?
show the reaction diagram
-
13% of the activity with quercetin, Co-QueD, 16% of the activity with quercetin, Ni-QueD
-
-
?
3,7,3',4'-tetrahydroxyflavone + O2
?
show the reaction diagram
-
35% of the activity with quercetin, Co-QueD, 15% of the activity with quercetin, Ni-QueD
-
-
?
fisetin + O2
?
show the reaction diagram
A7Y9J1, -
16.8% of the activity with quercetin
-
-
?
fisetin + O2
?
show the reaction diagram
A2VA43
i.e. 3,7,3',4'-tetrahydroxyflavone, 23% of the activity with quercetin
-
-
?
fisetin + O2
2-[[(3,4-dihydroxyphenyl)carbonyl]oxy]-4-hydroxybenzoate + CO
show the reaction diagram
Bacillus subtilis, Bacillus subtilis 168
-
1.22% activity compared to quercetin
-
-
?
galangin + O2
?
show the reaction diagram
A7Y9J1, -
21% of the activity with quercetin
-
-
?
galangin + O2
?
show the reaction diagram
A2VA43
i.e. 3,5,7-trihydroxyflavone, 28% of the activity with quercetin
-
-
?
galangin + O2
2,4-dihydroxy-6-[(phenylcarbonyl)oxy]benzoate + CO
show the reaction diagram
Bacillus subtilis, Bacillus subtilis 168
-
110% activity compared to quercetin
-
-
?
kaempferol + O2
?
show the reaction diagram
A7Y9J1, -
activity is 2.18fold higher than with quercetin
-
-
?
morin + O2
?
show the reaction diagram
A2VA43
i.e. 3,5,7,2',4'-pentahydroxyflavone, 1.7% of the activity with quercetin
-
-
?
myricetin + O2
?
show the reaction diagram
-
-
-
-
?
myricetin + O2
?
show the reaction diagram
A2VA43
i.e. 3,5,7,3',4',5'-hexahydroxyflavone. 49% of the activity with quercetin
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
A2VA43
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
low activity
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
100% activity
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
quercetin is a flavonol
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
Aspergillus flavus MTCC-2456
-
-, low activity
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
Bacillus subtilis 168
-
-, 100% activity
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
Aspergillus flavus MTCC-1783
-
-, low activity
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
Aspergillus flavus MTCC-2206
-
-, low activity
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
Aspergillus flavus PRL 1805
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
Aspergillus flavus MTCC-1884
-
-, low activity
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
Aspergillus flavus MTCC-1883
-
-, low activity
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
A2VA43
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
the mechanism consists in four successive steps, the first one concerns addition of O2 on the C2 carbon atom, the second corresponds to the closure of the endoperoxo intermediate. In the two last steps, bonds are broken to produce the depside and carbon monoxide. Addition of dioxygen on the C2 atom (step 1) is associated to a pyramidalization at the C2 carbon atom and to a rotation of the B-ring with respect to the conjugated A-C rings. The second step is the rate limiting one and the free energy barriers characterized for the four flavonoids are very close, reaching about 24 kcal/mol. Differences in the values are not significant enough to be exploited to rationalize the nonlinear evolution of the degradation rate. Moreover, the relatively high energy value is expected to be lowered by taking into account the whole environment
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
quercetin dioxygenase catalyzes the oxidation of the flavonol quercetin with dioxygen, cleaving the central heterocyclic ring and releasing CO
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO
show the reaction diagram
A7Y9J1, -
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO
show the reaction diagram
-
transcription of queD is triggered by quercetin and its 3-O-rhamnosylglucoside rutin, but not by the flavonol morin (3,5,7,2',4'-pentahydroxyflavone), the presumed quercetin degradation products protocatechuate and 2,4,6-trihydroxybenzoate or the sugars rhamnose and glucose. Quercetin-induced queD expression is not influenced by the presence of Ni(II)
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO
show the reaction diagram
-
-, transcription of queD is triggered by quercetin and its 3-O-rhamnosylglucoside rutin, but not by the flavonol morin (3,5,7,2',4'-pentahydroxyflavone), the presumed quercetin degradation products protocatechuate and 2,4,6-trihydroxybenzoate or the sugars rhamnose and glucose. Quercetin-induced queD expression is not influenced by the presence of Ni(II)
-
-
?
tamarixetin + O2
2,4-dihydroxy-6-[[(3-hydroxy-4-methoxyphenyl)carbonyl]oxy]benzoate + CO
show the reaction diagram
Bacillus subtilis, Bacillus subtilis 168
-
82.4% activity compared to quercetin
-
-
?
kaempferol + O2
?
show the reaction diagram
A2VA43
i.e. 3,5,7,4'-tetrahydroxyflavone. 70% of the activity with quercetin
-
-
?
additional information
?
-
-
high level of pirin leads to the resistance of poliovirus replication to quercetin by inactivating this flavonoid
-
-
-
additional information
?
-
A7Y9J1, -
flavonol, morin, 3,6-dihydroxyflavone and 3,7-dihydroxyflavone are transformed at a rate of less than 1% of that found for quercetin
-
-
-
additional information
?
-
A2VA43
no activity with luteolin
-
-
-
additional information
?
-
-
quercetin 2,3-dioxygenase is a copper-containing enzyme that catalyzes the insertion of molecular oxygen into polyphenolic flavonols
-
-
-
additional information
?
-
-
the enzyme opens up two C-C bonds of the heterocyclic ring of quercetin, a widespread plant flavonol
-
-
-
additional information
?
-
-
quercetin 2,3-dioxygenase activates molecular oxygen to catalyze the oxygenative ring-opening reaction of the O-heterocycle of quercetin to the corresponding depside (phenolic carboxylic acid esters) and carbon monoxide
-
-
-
additional information
?
-
-
the enzyme opens up two C-C bonds of the heterocyclic ring of quercetin, a widespread plant flavonol
-
-
-
additional information
?
-
A2VA43
no activity with luteolin
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
quercetin is a flavonol
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
quercetin dioxygenase catalyzes the oxidation of the flavonol quercetin with dioxygen, cleaving the central heterocyclic ring and releasing CO
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
Aspergillus flavus MTCC-2456
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
Bacillus subtilis 168
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
Aspergillus flavus MTCC-1783, Aspergillus flavus MTCC-2206
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
Aspergillus flavus PRL 1805
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
Aspergillus flavus MTCC-1884, Aspergillus flavus MTCC-1883
-
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO + H+
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-protocatechoylphloroglucinolcarboxylate + CO
show the reaction diagram
-
-
-
-
?
quercetin + O2
2-(3,4-dihydroxybenzoyloxy)-4,6-dihydroxybenzoate + CO
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
high level of pirin leads to the resistance of poliovirus replication to quercetin by inactivating this flavonoid
-
-
-
additional information
?
-
-
quercetin 2,3-dioxygenase is a copper-containing enzyme that catalyzes the insertion of molecular oxygen into polyphenolic flavonols
-
-
-
additional information
?
-
-
the enzyme opens up two C-C bonds of the heterocyclic ring of quercetin, a widespread plant flavonol
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
a non-heme redox metalloenzyme
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Co2+
-
Co2+ salt addition increases the activity of quercetin 2,3-dioxygenase 24fold. The Escherichia coli cultures were grown at 37C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM CoCl2, and allow to grow additional 4 h at 25C. The protein contains 0.65-0.8 atom of cobalt and 0.1 atom of iron per subunit.
Co2+
-
supplementing the cultures of strain FLA with CoCl2 results in 1.6fold higher quercetinase activity in crude extracts
Co2+
-
can partly substitute for Ni2+
Co2+
-
can partly substitute for Mn2+
copper
-
type II site
copper
-
single Cu(II) ion in active site
Cu
-
probably belongs to the nonblue class, two atoms per molecule of enzyme
Cu
-
cupric ion; probably belongs to the nonblue class, two atoms per molecule of enzyme
Cu
-
1-1.6 mol per mol enzyme, nonblue type 2 Cu2+ protein
Cu
-
0.8 mol per mol enzyme
Cu
A7Y9J1, -
contains 0.9 copper atoms per protein
Cu2+
-
Cu2+ salt addition increases the activity of quercetin 2,3-dioxygenase 1.4fold. The Escherichia coli cultures were grown at 37C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM CuCl2, and allow to grow additional 4 h at 25C.
Cu2+
-
required, the copper ion is mainly coordinated by three His residues and a water molecule in a distorted tetrahedral geometry. In a minor form, the metal is penta-coordinated by three His, a glutamate, and an aquo ligand in a trigonal bipyramidal geometry. The major role of the activesite metal ion could be to correctly position the substrate and to stabilize transition states and intermediates rather than to mediate electron transfer
Cu2+
-
required, the major role of the activesite metal ion could be to correctly position the substrate and to stabilize transition states and intermediates rather than to mediate electron transfer
Cu2+
-
required, mononuclear copper(II) active site, binding structure, X-ray diffraction and NMR analysis, overview. Direct coordinative interaction between copper(II) ion and the carboxylate group of Glu73. Complexes modeling, overview
Cu2+
-
can partly substitute for Mn2+
Cu2+
-
required, enzyme-bound, structure, overview. Manual docking, different geometries of the copper site
Fe2+
-
can partly substitute for Ni2+
Fe2+
-
can partly substitute for Mn2+
HNO
-
nitrosyl hydride replaces dioxygen in nitroxygenase activity of manganese quercetin dioxygenase resulting in the incorporation of both N and O atoms into the product. Turnover is demonstrated by consumption of quercetin and other related substrates under anaerobic conditions in the presence of HNO-releasing compounds and the enzyme. As with dioxygenase activity, a nonenzymatic base-catalyzed reaction of quercetin with HNO isobserved above pH 7, but no enhancement of this basal reactivity is found upon addition of divalent metal salts. Unique and regioselective N-containing products are characterized by MS analysis for both the enzymatic and nonenzymatic reactions
Iron
P42106
different coordination geometry in the two active sites of the dimer
Mn2+
-
Mn2+ salt addition increases the activity of quercetin 2,3-dioxygenase 35fold. The Escherichia coli cultures were grown at 37C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM MnSO4, and allow to grow additional 4 h at 25C. The protein containes 1.6-1.9 atoms of Mn/subunit.
Mn2+
-
required for activity
Mn2+
-
can partly substitute for Ni2+
Ni2+
-
Ni2+ salt addition increases the activity of quercetin 2,3-dioxygenase 2.6fold. The Escherichia coli cultures were grown at 37C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM NiCl2, and allow to grow additional 4 h at 25C.
Ni2+
-
supplementing the cultures of strain FLA with NiCl2 results in 6.1fold higher quercetinase activity in crude extracts
Ni2+
-
preferred divalent metal ion
Ni2+
-
can partly substitute for Mn2+. Nickel is a poor cofactor.
Mn2+
-
preferred divalent metal ion
additional information
-
Cd2+ does not increase the activity of quercetin 2,3-dioxygenase. The Escherichia coli cultures were grown at 37C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM CdCl2, and allow to grow additional 4 h at 25C.; Fe2+ does not increase the activity of quercetin 2,3-dioxygenase. The Escherichia coli cultures were grown at 37C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM FeCl2, and allow to grow additional 4 h at 25C.; Zn2+ does not increase the activity of quercetin 2,3-dioxygenase. The Escherichia coli cultures were grown at 37C and 200 rpm for 6 h, induced with isopropyl beta-D-thiogalactopyanoside to a final concentraton of 50 mg/l in the presence of 10 microM ZnSO4, and allow to grow additional 4 h at 25C.
additional information
-
no increase in activity is observed when Mn2+, Fe2+, Cu2+, or Zn2+ is added to the culture medium
additional information
-
fungal quercetinases appear to exclusively utilize a Cu2+ ion for catalysis
additional information
-
the bacterial enzyme is capable of using different divalent metal ions for catalysis, with preference Mn2+, Co2+, Fe2+, Ni2+, Cu2+in descending order, suggesting that the redox properties of the metal are relatively unimportant for the catalytic reaction. The major role of the active site metal ion could be to correctly position the substrate and to stabilize transition states and intermediates rather than to mediate electron transfer. The recombinant enzyme is able to exchange its active-site metal ion while retaining catalytic activity
additional information
-
fungal quercetinases appear to exclusively utilize a Cu2+ ion for catalysis
additional information
-
the bacterial enzyme is capable of using different divalent metal ions for catalysis, with preference Ni2+, Co2+, Mn2+, Fe2+ in descending order, suggesting that the redox properties of the metal are relatively unimportant for the catalytic reaction. The major role of the active site metal ion could be to correctly position the substrate and to stabilize transition states and intermediates rather than to mediate electron transfer
additional information
-
the enzyme from Bacillus subtilis is active with several divalent metal cofactors such as Fe, Mn, and Co, although Mn(II) is the preferred cofactor for this enzyme
additional information
-
a non-heme redox metalloenzyme
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(1Z,3Z)-N,N'-di(pyridin-2-yl)-1H-isoindole-1,3(2H)-diimine
-
-
1,10-phenanthroline
-
0.21 mM, 46% residual activity
1H-2-benzyl-3-hydroxy-4-oxoquinolin
A7Y9J1, -
competitive
2,2'-dipyridyl
-
0.18 mM, 57% residual activity
2-(4-{bis[1-methyl-2-(propan-2-yl)-1H-imidazol-4-yl]methyl}-1-methyl-1H-imidazol-2-yl)-2-methylpropanoic acid
-
-
2-mercaptoethanol
-
-
2-{4-[hydroxy(di-1H-imidazol-4-yl)methyl]-1H-imidazol-2-yl}-2-methylpropanoic acid
-
-
3-hydroxyflavone
-
competitive inhibition
8-hydroxyquinoline
-
at 1 mM or more
8-hydroxyquinoline
-
-
alpha-alpha'-dipyridyl
-
at 1 mM or more
alpha-naphthoquinoline
-
-
chloro[(4-phenoxy-1,4,7-triazonan-1-yl-kappa3N1,N4,N7)acetato-kappaO]copper
-
-
diethyldithiocarbamate
-
at microM concentrations or less
diethyldithiocarbamate
-
-
diethyldithiocarbamate
-
-
diethyldithiocarbamate
-
-
diethyldithiocarbamate
-
89 mM, 62% residual activity
diethyldithiocarbamate
-
Co QueD: 90% activity remains after 15 min incubation with 1 mM reagent, Mn QueD: 84% activity remains after 15 min incubation with 1 mM reagent
diethyldithiocarbamate
A7Y9J1, -
100 nM, complete inhibition
Diphenylthiocarbazone
-
-
Dithiol
-
at microM concentrations or less
dithiothreitol
-
-
Dithizone
-
at microM concentrations or less
DTT
-
Mn QueD: 96% activity remains after 15 min incubation with 1 mM reagent
EDTA
-
94 mM, 94% residual activity
EDTA
-
Mn QueD: 90% activity remains after 15 min incubation with 1 mM reagent
ethylxanthate
-
competitive inhibition, at microM concentrations or less
H2O2
-
Co QueD: 93% activity remains after 15 min incubation with 1 mM reagent, Mn QueD: 82% activity remains after 15 min incubation with 1 mM reagent
Kojic acid
-
5 mM, 50% residual activity
Kojic acid
-
Co QueD: 92% activity remains after 15 min incubation with 1 mM reagent, Mn QueD: 100% activity remains after 15 min incubation with 1 mM reagent
morin
-
competitive inhibition
NaCN
-
Co QueD: 91% activity remains after 15 min incubation with 1 mM reagent, Mn QueD: 89% activity remains after 15 min incubation with 1 mM reagent
O-ethylxanthate
-
6 mM, 51% residual activity
O-ethylxanthic acid
-
Co QueD: 69% activity remains after 15 min incubation with 1 mM reagent, Mn QueD: 100% activity remains after 15 min incubation with 1 mM reagent
o-phenanthroline
-
at 1 mM or more
Sodium ascorbate
-
Mn QueD: 80% activity remains after 15 min incubation with 1 mM reagent
Sodium dithionite
-
-
Toluene-3,4-dithiol
-
-
[3-(hydroxy-kappaO)-2-phenyl-4H-chromen-4-onato-kappaO4][bis(triphenylphosphane)]copper
-
-
{N,N-bis[(1-methyl-1H-imidazol-4-yl-kappaN3)methyl]glycinato-kappa2N,O}(dichloro)ferrate(1-)
-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
DTT
-
Co QueD: 110% activity remains after 15 min incubation with 1 mM reagent
EDTA
-
Co QueD: 110% activity remains after 15 min incubation with 1 mM reagent
NaN3
-
Co QueD: 110% activity remains after 15 min incubation with 1 mM reagent, Mn QueD = 100% activity remains after 15 min incubation with 1 mM reagent
Sodium ascorbate
-
Co QueD: 110% activity remains after 15 min incubation with 1 mM reagent
Tiron
-
0.8 mM, 25% activation
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.085
-
fisetin
A7Y9J1, -
pH 6, 25C
0.02
-
galangin
A7Y9J1, -
pH 6, 25C
0.079
-
oxygen
-
Co quercetin 2,3-dioxygenase (0.65 atoms Co/subunit)
0.09
-
oxygen
-
Mn quercetin 2,3-dioxygenase (1.8 atoms Mn/subunit)
0.12
-
oxygen
-
-
0.15
-
oxygen
-
Fe quercetin 2,3-dioxygenase
0.0008
-
quercetin
-
pH 7.5, 25C
0.00096
-
quercetin
-
recombinant Co-QueD, 50 mM Tris-HCl buffer pH 8
0.004
-
quercetin
-
Mn quercetin 2,3-dioxygenase (1.8 atoms Mn/subunit)
0.004
-
quercetin
-
pH 7.0, temperature not specified in the publication
0.0052
-
quercetin
-
-
0.0052
-
quercetin
-
Fe quercetin 2,3-dioxygenase
0.00575
-
quercetin
-
recombinant Ni-QueD, 50 mM Tris-HCl buffer pH 8
0.0066
-
quercetin
-
-
0.0075
-
quercetin
-
Co quercetin 2,3-dioxygenase (0.65 atoms Co/subunit)
0.0125
-
quercetin
-
-
0.013
-
quercetin
-
-
0.014
-
quercetin
-
-
0.0141
-
quercetin
-
-
0.019
-
quercetin
A7Y9J1, -
pH 6, 25C
0.013
-
kaempferol
A7Y9J1, -
pH 6, 25C
additional information
-
additional information
-
kinetics of enzyme-substrate complexes of the active site of MII-containing quercetin 2,3-dioxygenase, overview
-
additional information
-
additional information
-
kinetics, modeling, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
56.7
-
fisetin
A7Y9J1, -
pH 6, 25C
33.3
-
galangin
A7Y9J1, -
pH 6, 25C
466.7
-
kaempferol
A7Y9J1, -
pH 6, 25C
12.5
-
O2
-
recombinant Co-QueD, 50 mM Tris-HCl buffer pH 8
19
-
O2
-
recombinant Ni-QueD, 50 mM Tris-HCl buffer pH 8
0.65
-
quercetin
-
Fe quercetin 2,3-dioxygenase
1.45
-
quercetin
-
-
6.7
-
quercetin
-
Co quercetin 2,3-dioxygenase (0.65 atoms Co/subunit)
7.6
-
quercetin
-
recombinant Co-QueD, 50 mM Tris-HCl buffer pH 8
25
-
quercetin
-
Mn quercetin 2,3-dioxygenase (1.8 atoms Mn/subunit)
40.1
-
quercetin
-
recombinant Ni-QueD, 50 mM Tris-HCl buffer pH 8
166.7
-
quercetin
A7Y9J1, -
pH 6, 25C
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
33000
-
quercetin
-
pH 7.0, temperature not specified in the publication
16073
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.004
-
1H-2-benzyl-3-hydroxy-4-oxoquinolin
A7Y9J1, -
pH 6, 25C
0.034
-
8-hydroxyquinoline
-
-
0.745
-
alpha-alpha'-dipyridyl
-
-
0.11
-
alpha-naphthoquinoline
-
-
0.00027
-
ethylxanthate
-
-
0.115
-
o-phenanthroline
-
-
1.55
-
Quinoline
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.0012
-
-
native enzyme, using quercetin as substrate, at 25C, pH 7.5
0.0276
-
-
recombinant enzyme, using fisetin as substrate, at 25C, pH 7.5
1.5
-
-
25C, pH 7.5
1.87
-
-
recombinant enzyme, using tamarixetin as substrate, at 25C, pH 7.5
2.27
-
-
recombinant enzyme, using quercetin as substrate, at 25C, pH 7.5
2.49
-
-
recombinant enzyme, using galangin as substrate, at 25C, pH 7.5
4
-
-
Fe-QueD
28
-
-
Co-QueD
144
-
-
Ni-QueD
175
-
A7Y9J1, -
-
additional information
-
-
high specific QueD activity is found in crude cell extracts when the growth medium is supplemented with NiCl2 or CoCl2, but not when Mn2+, Fe2+, Cu2+, or Zn2+ is added, the activity of the purified QueD proteins depends strongly on the nature of the metal ion cofactor
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
-
A7Y9J1, -
0.1 M MES buffer
7
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2
6
-
Britton-Robinson buffer, composed of H3PO4, acetic acid, H3BO3
5.5
6.7
-
50 mM MES buffer
6
9
-
50 mM MES-Tris buffer
additional information
-
-
pH profiles are determined spectrophotometrically on a Hp 8453 photodiode array spectrophotometer
additional information
-
-
nonenzymatic, basal reactivity of sodium trioxodinitrate/Na2N2O3 (Angeli's salt) as HNO donor with quercetin initiates above pH 7.0
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
40
-
A7Y9J1, -
quercetin oxidation
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.9
-
-
calculated
additional information
-
-
pKa determined from the Vmax profile is 5.6. The pKa determined from the Vmax/KM pH profile is 5.8.
additional information
-
A7Y9J1, -
isoelectric focusing analysis of the purified enzyme reveals some heterogeneity since two major bands with apparent pI of 5.81 and 6.07 as well as two minor bands with apparent pI of 5.55 and 6.38 are observed. The purified enzyme is a mixture of at least four glycoforms
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
HeLa cells contain higher content of pirin protein than normal kidney human epithelial or HEK-293 cells
Manually annotated by BRENDA team
-
HeLa cells contain higher content of pirin protein than normal kidney human epithelial or HEK-293 cells
Manually annotated by BRENDA team
-
HeLa cells contain higher content of pirin protein than normal kidney epithelial or HEK-293 cells
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Aspergillus flavus MTCC-1783, Aspergillus flavus MTCC-1883, Aspergillus flavus MTCC-1884, Aspergillus flavus MTCC-2206, Aspergillus flavus MTCC-2456, Aspergillus flavus PRL 1805
-
-
-
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
23040
-
-
calculated value of the hexahistidine-tagged QueD monomer
40000
-
-
about 40000 Da, SDS-PAGE
45000
-
-
SDS-PAGE
55100
-
-
SDS-PAGE
56000
-
A7Y9J1, -
gel filtration
63000
-
-
gel filtration
65000
-
-
gel permeation chromatography, native Ni-QueD
65100
-
-
gel permeation chromatography, native Co-QueD
110000
-
-
-
110000
-
-
low speed sedimentation
111400
-
-
gel filtration
135000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 37640, MALDI-TOF, SDS-PAGE
?
-
x * 23041, calculated from sequence
?
-
x * 23041, calculated from sequence
-
dimer
P42106
crystallization data
dimer
-
SDS-PAGE
dimer
-
SDS-PAGE
-
homodimer
-
bicupin domain structure
homodimer
-
monocupin domain structure
homodimer
-
bicupin enzyme
homodimer
-
he enzyme forms homodimers, which are stabilized by an N-linked heptasaccharide at the dimer interface. The mononuclear type 2 copper center displays two distinct geometries: a distorted tetrahedral coordination, formed by His66, His68, His112, and a water molecule, and a distorted trigonal bipyramidal environment, which additionally comprises Glu73. Manual docking of the substrate quercetin into the active site showed that the different geometries of the copper site might be of catalytic importance
homodimer
-
monocupin domain structure
-
monomer
A7Y9J1, -
1 * 55000, SDS-PAGE
trimer
-
alpha 1 * 69574 + beta 1 * 51733 + gamma 1 * 26510, MALDI-TOF
trimer
-
3 * 23041, gel permeation chromatography
trimer
-
3 * 23041, gel permeation chromatography
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
containing 27.5% carbohydrate
glycoprotein
-
-
glycoprotein
Aspergillus flavus PRL 1805
-
containing 27.5% carbohydrate
-
glycoprotein
-
-
glycoprotein
-
N-linked oligo-mannose type glycan chains, 46-54% carbohydrate
glycoprotein
A7Y9J1, -
the purified enzyme is a mixture of at least four glycoforms
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
copper-containing quercetin 2,3-dioxygenase, X-ray diffraction structure determination and analysis at 1.6 A resolution
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
-
-
precipitates below
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
storage in buffer, loss of 25% of activity during 48 h, high concentrations of ammonium chloride stabilize, ascorbic acid and cysteine do not
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C, stable for several weeks
-
stable at -20C
-
4C, 6 months, the enzyme retains 81% of its initial activity
A7Y9J1, -
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
DEAE-Sephacel column equilibrated with 50 mM Tris.HCl, pH 7.5, and eluted with a NaCl gradient (0-600 mM). Ultrogel ACA 34 column, eluted with 50 mM Tris.HCl, pH 7.5, and 100 mM Nacl. DEAE-Sepharose column equilibrated with 50 mM Tris.HCl, pH 7,5, eluted with a gradient of NaCl (100-500 mM).
-
partial purification by ammonium sulfate precipitation and DEAE Toyopearl column chromatography
-
metal chelate affinity chromatography
-
recombinant QueD protein with a C-terminal hexahistidine-tag
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Aspergillus awamori
-
expressed in Escherichia coli BL21(DE3)
-
expressed in Escherichia coli BL21(DE3) cells
-
expression in Escherichia coli
-
expression of Fe-QDO in Escherichia coli
-
expression in Escherichia coli BL21(DE3) (pLysS, pET23a-queD) as a His6-tagged protein
-
overexpression in Escherichia coli or in in Streptomyces lividans TK23
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
E76A
-
site-directed mutagenesis of Ni-QueD, exhibits marginal quercetinase activity of 0.014 unit/mg, corresponding to a more than 10000fold decrease in specific activity, Glu76 of QueD is part of the strictly conserved cupin motif and thus is assumed to be a ligand to the metal center
E76A
-
site-directed mutagenesis of Ni-QueD, exhibits marginal quercetinase activity of 0.014 unit/mg, corresponding to a more than 10000fold decrease in specific activity, Glu76 of QueD is part of the strictly conserved cupin motif and thus is assumed to be a ligand to the metal center
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
synthesis
-
optimization of enzyme production by variation of rutin concentration, nitrogen source and concentration, salt and metal salt concentration, yeast extract concentration and pH value. six-fold improvement of enzyme activity reaching a maximum activity of 0.000708 mM per min and ml of culture supernatant