Information on EC 2.4.1.91 - flavonol 3-O-glucosyltransferase

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The expected taxonomic range for this enzyme is: Magnoliophyta

EC NUMBER
COMMENTARY hide
2.4.1.91
-
RECOMMENDED NAME
GeneOntology No.
flavonol 3-O-glucosyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
UDP-glucose + a flavonol = UDP + a flavonol 3-O-beta-D-glucoside
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexosyl group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
-
daphnetin modification
-
-
Flavone and flavonol biosynthesis
-
-
flavonol acylglucoside biosynthesis I - kaempferol derivatives
-
-
flavonol acylglucoside biosynthesis III - quercetin derivatives
-
-
kaempferol gentiobioside biosynthesis
-
-
kaempferol glycoside biosynthesis (Arabidopsis)
-
-
kaempferol triglucoside biosynthesis
-
-
Metabolic pathways
-
-
myricetin gentiobioside biosynthesis
-
-
quercetin gentiotetraside biosynthesis
-
-
quercetin glucoside biosynthesis (Allium)
-
-
quercetin glycoside biosynthesis (Arabidopsis)
-
-
quercetin triglucoside biosynthesis
-
-
rutin biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
UDP-glucose:flavonol 3-O-D-glucosyltransferase
Acts on a variety of flavonols, including quercetin and quercetin 7-O-glucoside. Different from EC 2.4.1.81 (flavone 7-O-beta-glucosyltransferase).
CAS REGISTRY NUMBER
COMMENTARY hide
50812-18-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
cv. Red Danish
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
f. Ciliato-dentatus
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
cultivar Nuomici
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
var. Mairei
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
cultivars Luchia Lady, Aphrodite, Wedding Promenade, and I-Hsin Lucky Girl x I-Hsin Song
-
-
Manually annotated by BRENDA team
Matsum
-
-
Manually annotated by BRENDA team
Matsum
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
cv. Apeldoorn
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
Kadainou R-1, interspecific hybrid grape between Vitis ficifolia variant Ganebu and Vitis vinifera cultivar Muscat of Alexandria
UniProt
Manually annotated by BRENDA team
cultivar Concord
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
enzyme-suppressed Phalaenopsis exhibit various levels of flower color fading that is well correlated with the extent of reduced level of transcriptional activity. Furthermore, there is a significant decrease in anthocyanin content in the enzyme-suppressed Phalaenopsis flowers
metabolism
-
in anthocyanin biosynthesis, UDP-glucose: anthocyanidin 3-O-glucosyltransferase catalyzes the transfer of the glucosyl moiety from UDP-glucose to the 3-hydroxyl group of anthocyanidins, producing the first stable anthocyanins
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
dTDP-glucose + quercetin
dTDP + quercetin 3-O-beta-D-glucoside
show the reaction diagram
-
-
-
-
?
dTDP-xylose + quercetin
dTDP + quercetin 3-O-beta-D-xylose
show the reaction diagram
-
low activity
-
-
?
GDP-glucose + quercetin
GDP + quercetin 3-O-beta-D-glucoside
show the reaction diagram
-
low activity
-
-
?
UDP-5-thio-glucose + quercetin
UDP + quercetin 3-O-5-thio-beta-D-glucoside
show the reaction diagram
-
low activity
-
-
?
UDP-alpha-D-glucose + 1-naphthol
UDP + naphthalen-1-yl beta-D-glucopyranoside
show the reaction diagram
-
glucosylated by enzyme FaGT6 and FaGT7
-
-
?
UDP-alpha-D-glucose + 2-naphthol
UDP + naphthalen-2-yl beta-D-glucopyranoside
show the reaction diagram
-
glucosylated by enzyme FaGT6 and FaGT7
-
-
?
UDP-alpha-D-glucose + 3,7-dihdroxyflavone
UDP + 3-[(beta-D-glucopyranosyl)oxy]-7-hydroxyflavone
show the reaction diagram
-
glucosylated by enzyme FaGT6 and FaGT7
-
-
?
UDP-alpha-D-glucose + 3-hydroxycoumarin
UDP + 3-[(beta-D-glucopyranosyl)oxy]-coumarin
show the reaction diagram
-
glucosylated by enzyme FaGT6 and FaGT7
-
-
?
UDP-alpha-D-glucose + 3-hydroxyflavone
UDP + 3-[(beta-D-glucopyranosyl)oxy]-flavone
show the reaction diagram
UDP-alpha-D-glucose + 6-hydroxycoumarin
UDP + 6-[(beta-D-glucopyranosyl)oxy]-coumarin
show the reaction diagram
-
glucosylated by enzyme FaGT6
-
-
-
UDP-alpha-D-glucose + 7-hydroxycoumarin
UDP + 7-[(beta-D-glucopyranosyl)oxy]-coumarin
show the reaction diagram
-
glucosylated by enzyme FaGT6 and FaGT7
-
-
?
UDP-alpha-D-glucose + cyanidin
UDP + cyanidin 3-O-beta-D-glucoside
show the reaction diagram
UDP-alpha-D-glucose + daphnetin
UDP + ?
show the reaction diagram
-
glucosylated by enzyme FaGT6 and FaGT7
-
-
?
UDP-alpha-D-glucose + delphinidin
UDP + delphinidin 3-O-beta-D-glucoside
show the reaction diagram
UDP-alpha-D-glucose + dihydroquercetin
UDP + quercetin 3-O-beta-D-glucoside
show the reaction diagram
weak substrate
-
-
?
UDP-alpha-D-glucose + galangin
UDP + galangin 3-O-beta-D-glucoside
show the reaction diagram
-
glucosylated by enzyme FaGT6 and FaGT7
-
-
?
UDP-alpha-D-glucose + isorhamnetin
UDP + isorhamnetin 3-O-beta-D-glucoside
show the reaction diagram
UDP-alpha-D-glucose + kaempferol
UDP + kaempferol 3-O-beta-D-glucoside
show the reaction diagram
UDP-alpha-D-glucose + malvidin
UDP + malvidin 3-O-beta-D-glucoside
show the reaction diagram
UDP-alpha-D-glucose + morin
UDP + morin 3-O-beta-D-glucoside
show the reaction diagram
-
glucosylated by enzyme FaGT6
-
-
?
UDP-alpha-D-glucose + myricetin
UDP + myricetin 3-O-beta-D-glucoside
show the reaction diagram
UDP-alpha-D-glucose + pelargonidin
UDP + pelargonidin 3-O-beta-D-glucoside
show the reaction diagram
UDP-alpha-D-glucose + peonidin
UDP + peonidin 3-O-beta-D-glucoside
show the reaction diagram
UDP-alpha-D-glucose + petunidin
UDP + petunidin 3-O-beta-D-glucoside
show the reaction diagram
UDP-alpha-D-glucose + quercetin
UDP + quercetin 3-O-beta-D-glucoside
show the reaction diagram
UDP-alpha-D-glucose + scopoletin
UDP + ?
show the reaction diagram
-
glucosylated by enzyme FaGT6 and FaGT7
-
-
?
UDP-alpha-D-glucose + taxifolin
UDP + taxifolin 3-O-beta-D-glucoside
show the reaction diagram
-
glucosylated by enzyme FaGT7
-
-
?
UDP-galactose + cyanidin
?
show the reaction diagram
galactose can be transferred from UDP-galactose to the 3-position of cyanidin with low specificity relative to the analogous glucosyltransferase reactions
-
-
?
UDP-galactose + delphinidin
?
show the reaction diagram
galactose can be transferred from UDP-galactose to the 3-position of delphinidin with low specificity relative to the analogous glucosyltransferase reactions
-
-
?
UDP-galactose + quercetin
UDP + quercetin 3-O-beta-D-galactoside
show the reaction diagram
-
low activity
-
-
?
UDP-glucose + 2',4'-dihydroxyflavone
UDP + ?
show the reaction diagram
-
8.3% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + 2'-hydroxyflavone
UDP + 2'-hydroxyflavone 2'-beta-D-glucoside
show the reaction diagram
-
15% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + 2,3,6-trichlorophenol
UDP + 2,3,6-trichlorophenylglucoside
show the reaction diagram
-
9% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + 2,4,5-trichlorophenol
UDP + 2,4,5-trichlorophenylglucoside
show the reaction diagram
-
-
-
-
?
UDP-glucose + 2,4,6-trichlorophenol
UDP + 2,4,6-trichlorophenylglucoside
show the reaction diagram
-
4% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + 2,4-dinitrophenol
UDP + 2,4-dinitrophenylglucoside
show the reaction diagram
-
2.4% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + 4'-hydroxyflavone
UDP + 4'-hydroxyflavone 4'-beta-D-glucoside
show the reaction diagram
-
4% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + 4-nitrophenol
UDP + 4-nitrophenylglucoside
show the reaction diagram
-
23% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + 5-hydroxyflavone
UDP + 5-hydroxyflavone 5-beta-D-glucoside
show the reaction diagram
-
2.7% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + 6-hydroxyflavone
UDP + 6-hydroxyflavone 6-beta-D-glucoside
show the reaction diagram
-
24% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + 7-hydroxyflavone
UDP + 7-hydroxyflavone 7-beta-D-glucoside
show the reaction diagram
-
4.8% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + apigenin
UDP + apigenin 3-O-glucoside
show the reaction diagram
-
8.7% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + coumestrol
UDP + coumestrol 3-O-glucoside
show the reaction diagram
-
33% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + delphinidin
UDP + delphinidin 3-O-glucoside
show the reaction diagram
-
-
-
-
?
UDP-glucose + esculetin
UDP + ?
show the reaction diagram
-
glucosylated by enzyme FaGT6 and FaGT7
-
-
?
UDP-glucose + genistein
UDP + ?
show the reaction diagram
-
7.4% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + isorhamnetin
UDP + isorhamnetin 3-O-beta-D-glucoside
show the reaction diagram
-
-
-
-
?
UDP-glucose + isosakuranetin
UDP + ?
show the reaction diagram
-
at 11.2% the activity with kaempferol
-
-
?
UDP-glucose + luteolin
UDP + ?
show the reaction diagram
UDP-glucose + myricetin
UDP + myricetin 3-O-beta-D-glucoside
show the reaction diagram
40.7% activity compared to quercetin, glucosylation occurs at the 3 hydroxyl position
-
-
?
UDP-glucose + pelargonidin
UDP + pelargonidin 3-O-glucoside
show the reaction diagram
-
at 4% the activity with kaempferol
-
-
?
UDP-glucose + pentachlorophenol
UDP + pentachlorophenylglucoside
show the reaction diagram
-
4% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + phenol
UDP + phenyglucoside
show the reaction diagram
-
2.4% of the activity with 2,4,5-trichlorophenol
-
-
?
UDP-glucose + quercetin
UDP + quercetin 3-O-beta-D-glucoside
show the reaction diagram
-
-
-
-
?
UDP-mannose + quercetin
UDP + quercetin 3-O-beta-D-mannoside
show the reaction diagram
-
very low activity
-
-
?
UDP-N-acetylglucose + quercetin
UDP + quercetin 3-O-N-acetylglucoside
show the reaction diagram
-
low activity
-
-
?
UDP-xylose + quercetin
UDP + quercetin 3-O-beta-D-xylose
show the reaction diagram
-
low activity
-
-
?
UDPgalactose + isorhamnetin
UDP + isorhamnetin 3-O-glucoside
show the reaction diagram
-
isorhamnetin is the best acceptor, 45% of the activity compared to reaction with UDPglucose
-
-
r
UDPglucose + 3'-O-methylquercetin
UDP + 3'-O-methylquercetin 3-O-glucoside
show the reaction diagram
-
28% of the activity with quercetin
-
-
?
UDPglucose + 3,5,7-trihydroxyflavanone
UDP + 3,5,7-trihydroxyflavanone 3-O-glucoside
show the reaction diagram
-
isoenzyme F3GT1: 22% of the activity with kaempferol, isoenzyme F3GT2: 12% of the activity with kaempferol
-
-
?
UDPglucose + 4,7-dihydroxyflavonol
UDP + 4,7-dihydroxyflavonol 3-O-glucoside
show the reaction diagram
-
27% of the activity with quercetin
-
-
?
UDPglucose + 5-deoxyquercetin
UDP + 5-deoxyquercetin 3-O-glucoside
show the reaction diagram
-
57% of the activity with quercetin
-
-
?
UDPglucose + 7,4'-dihydroxyflavonol
UDP + 7,4'-dihydroxyflavonol 3-O-glucoside
show the reaction diagram
-
isoenzyme F3GT1: 33% of the activity with kaempferol, isoenzyme F3GT2: 34% of the activity with kaempferol
-
-
?
UDPglucose + a flavonol
UDP + a flavonol 3-O-D-glucoside
show the reaction diagram
UDPglucose + apigenin
UDP + apigenin 3-O-glucoside
show the reaction diagram
-
at 4.2% the activity with kaempferol
-
-
?
UDPglucose + cyanidin
UDP + cyanidin 3-O-glucoside
show the reaction diagram
UDPglucose + dihydrokaempferol
UDP + dihydrokaempferol 3-O-glucoside
show the reaction diagram
UDPglucose + dihydroquercetin
UDP + dihydroquercetin 3-O-glucoside
show the reaction diagram
UDPglucose + fisetin
UDP + fisetin 3-O-glucoside
show the reaction diagram
UDPglucose + kaempferol
UDP + kaempferol 3-O-glucoside
show the reaction diagram
UDPglucose + kaempferol 5,7,4'-trimethyl ether
UDP + kaempferol 5,7,4'-trimethyl ether 3-O-glucoside
show the reaction diagram
UDPglucose + kaempferol 7-O-glucoside
UDP + kaempferol 3,7-O-diglucoside
show the reaction diagram
UDPglucose + kaempferol-4'-O-methylether
UDP + ?
show the reaction diagram
UDPglucose + malvidin
UDP + malvidin 3-O-glucoside
show the reaction diagram
UDPglucose + myricetin
UDP + myrecetin 3-O-glucoside
show the reaction diagram
UDPglucose + naringenin
UDP + naringenin 3-O-glucoside
show the reaction diagram
-
at 3.2% the activity with kaempferol
-
-
?
UDPglucose + quercetin
UDP + quercetin 3-O-glucoside
show the reaction diagram
UDPglucose + quercetin 5-methyl ether
UDP + quercetin 5-methyl ether 3-O-glucoside
show the reaction diagram
UDPglucose + quercetin 7-O-glucoside
UDP + quercetin 3,7-O-diglucoside
show the reaction diagram
UDPglucose + rhamnetin
UDP + rhamnetin 3-O-glucoside
show the reaction diagram
UDPglucose + rutin
UDP + rutin 3-O-glucoside
show the reaction diagram
-
4% of the activity with isorhamnetin
-
-
r
additional information
?
-
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
UDP-alpha-D-glucose + cyanidin
UDP + cyanidin 3-O-beta-D-glucoside
show the reaction diagram
-
-
-
-
?
UDP-alpha-D-glucose + isorhamnetin
UDP + isorhamnetin 3-O-beta-D-glucoside
show the reaction diagram
-
glucosylated by enzyme FaGT6 and FaGT7
-
-
?
UDP-alpha-D-glucose + kaempferol
UDP + kaempferol 3-O-beta-D-glucoside
show the reaction diagram
UDP-alpha-D-glucose + pelargonidin
UDP + pelargonidin 3-O-beta-D-glucoside
show the reaction diagram
-
-
-
-
?
UDP-alpha-D-glucose + quercetin
UDP + quercetin 3-O-beta-D-glucoside
show the reaction diagram
UDP-glucose + isorhamnetin
UDP + isorhamnetin 3-O-beta-D-glucoside
show the reaction diagram
-
-
-
-
?
UDPglucose + a flavonol
UDP + a flavonol 3-O-D-glucoside
show the reaction diagram
additional information
?
-
-
the enzyme is involved in flavonoid biosynthesis, overview
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
UDP
-
dependent
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Na2SO4
118% relative activity at 1 mM
Succinic anhydride
115% relative activity at 1 mM
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,3-Butanedione
46% residual activity at 10 mM
cyanidin
the recombinant enzyme exhibits pronounced substrate inhibition by cyanidin at 0.1 mM
dithioerythritol
EDTA
-
10 mM, slight inhibition
Fe2+
0.09% residual activity at 10 mM
iodoacetamide
iodoacetate
N-bromo-succinimide
17% residual activity at 10 mM
Phenylmercuriacetate
Tetranitromethane
13% residual activity at 10 mM
Tris
Tris inhibits flavonol 3-O-GT as activity levels are significantly lower than corresponding buffers across its entire buffer range
UDP
6% residual activity at 10 mM
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
dithioerythritol
glutathione
-
1 mM, activation to 118%
glycine-HCl
-
glycine-HCl buffer stimulates about 2fold at pH 9 in comparison with Tris-HCl buffer
Sucrose
-
5%, activation to 123%
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0106
2,4,5-Trichlorophenol
-
-
0.0053
3-hydroxyflavone
-
pH 7.0, 30C, enzyme FaGT6
0.00479 - 0.07
cyanidin
0.016
delphinidin
-
recombinant enzyme
0.27
fisetin
-
-
0.0035 - 0.2
isorhamnetin
0.00067 - 0.69
kaempferol
0.0357
malvidin
-
recombinant enzyme
0.033
myricetin
apparent value, in 50 mM phosphate buffer (pH 7.5), at 40C
0.001 - 0.126
quercetin
0.6 - 2
UDP-alpha-D-glucose
1.67
UDP-D-glucose
-
-
0.0124 - 0.669
UDP-glucose
0.0098 - 1.9
UDPglucose
additional information
additional information
-
kinetics, recombinant enzyme
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000702
cyanidin
Vitis labrusca
A6N928
in 100 mM Tris-HCl pH 8.0, at 30C
0.5955 - 1.25
kaempferol
0.6252
myricetin
Citrus x paradisi
C5MR71
apparent value, in 50 mM phosphate buffer (pH 7.5), at 40C
0.000113 - 1.047
quercetin
0.000247
UDP-alpha-D-glucose
Vitis labrusca
A6N928
using cyanidin as cosubstrate, in 100 mM Tris-HCl pH 8.0, at 30C
1.442
UDP-glucose
Citrus x paradisi
C5MR71
apparent value, in 50 mM phosphate buffer (pH 7.5), at 40C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.146
cyanidin
Vitis labrusca
A6N928
in 100 mM Tris-HCl pH 8.0, at 30C
1072
49.6
kaempferol
Citrus x paradisi
C5MR71
apparent value, in 50 mM phosphate buffer (pH 7.5), at 40C
408
18.9
myricetin
Citrus x paradisi
C5MR71
apparent value, in 50 mM phosphate buffer (pH 7.5), at 40C
484
0.052 - 15.6
quercetin
0.00027
UDP-alpha-D-glucose
Vitis labrusca
A6N928
using cyanidin as cosubstrate, in 100 mM Tris-HCl pH 8.0, at 30C
364
2.2
UDP-glucose
Citrus x paradisi
C5MR71
apparent value, in 50 mM phosphate buffer (pH 7.5), at 40C
64
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000716
-
activity with 2,4,5-trichlorophenol
0.00143
-
activity with quercetin
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.8 - 6.2
-
-
7
-
assay at
7.2 - 7.3
-
assay at
7.8
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 9
-
pH 4.0: about 60% of maximal activity, pH 9.0: about 70% of maximal activity
4.5 - 8
-
pH 4.5: about 80% of maximal activity, pH 8.0: about 55% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
28
-
assay at
37
-
assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
isoelectric focusing
5.2
calculated from amino acid sequence
5.6
-
calculated from amino acid sequence
6.3
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
highest expression
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40000
-
gel filtration
48000
-
gel filtration
49000
-
gel filtration
50190
calculated from amino acid sequence
51200
calculated from amino acid sequence
59000
-
gel filtration
66000
-
SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
the structure of UGT71G1 consists of two N- and C-terminal domains with similar Rossmann-type folds and belongs to the GT-B fold, the N-terminal domain contains a central seven-stranded parallel beta sheets flanked by eight alpha helices on both sides and a small two stranded beta sheets, the C-terminal domain contains a six stranded beta sheet flanked by eight alpha helices, the two domains pack very tightly and form a deep cleft with a UDP molecule bound, structure comparisons, overview
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
wild-type and selenomethionine-labeled enzyme in complex with UDP and UDP-glucose, hanging drop vapour diffusion method, 5 mg/ml protein is mixed with 5 mM UDP-galactose and 5 mM quercetin at a 2:1 v/v ratio, mixed with an equal volume of reservoir solution containing 40% w/v PEG 3350, 0.2 M ammonium acetate, and 0.1 M sodium citrate, pH 5.6, equilibration over the reservoir solution at 20C, 2-5 days, X-ray diffraction structure determination and analysis at 2.0-2.6 A resolution, molecular docking
purified recombinant detagged enzyme bound to UDP-2-fluoroglucose and kaempferol, or with UDP and quercetin, hanging drop method, 4.8 mg/ml protein in 5 mM Tris (2-carboxyethyl) phosphine HCl, 10 mM TrisCl, pH 8.3, diluted 1:1 with mother liquor which contains 1822% polyethylene glycol 10 000, 0.1M Bis-Tris propane, pH 7.0, and 00.5% v/v Pluronic F-68, crystals appear within 24 h, X-ray diffraction structure determination and analysis at 2.2 A resolution
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8.5
reduction to 59% maximal activity at pH 5.5 and 57% at pH 8.5
706126
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40 - 60
the enzyme is stable when preincubated for up to 10 min at 40C with a significant decrease in activity at 50C and an almost complete loss of detectable activity at 60C and beyond
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
activities recorded in phosphate, MOPS, and bicine buffers are not significantly different, but assays in Tris buffer result in a reduction of enzyme activity by 51% under optimal pH conditions
at least three freeze-thaw cycles can be tolerated without apparent loss of activity
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C or 4C, the recombinant enzyme is stable for at least 120 h
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-20C, 20 mM imidazole-HCl, pH 8.0, 14 mM 2-mercaptoethanol, 10% glycerol, 35% loss of activity after 2 days, 61% loss of activity after 1 week
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-20C, 20 mM Tris-HCl, pH 7.5, 14 mM 2-mercaptoethanol, 10% glycerol, 15% loss of activity after 2 days
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-80C, stable for many months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; partial
-
affinity-purified on immobilized reduced glutathione resin
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glutathione Sepharose column chromatography
isoenzyme F3GT1 and F3GT2
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native enzyme from fruits, partially
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partial
recombinant enzyme
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recombinant His-tagged enzyme from Escherichia coli to homogeneity, the His-tag is cleaved off
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recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, anion exchange chromatography, and gel filtration
Talon metal affinity column chromatography
TALON metal affinity resin column chromatography, gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a binary vector containing an UDPglucose:flavonoid-3-O-glucosyltransferase cDNA under the control of the cauliflower mosaic virus 35S promoter is used to transform Eustoma grandiflorum Grise. Of four independent transgenic lines recovered, one produces high levels of the UDPglucose:flavonoid-3-O-glucosyltransferase transcript and synthesizes 3-O-glucosylated anthocyanins novel to Eusoma grandiflorum, as well as enhanced levels of 3-O-glucosylated flavonols
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expressed in Escherichia coli
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expressed in Escherichia coli BL21
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expressed in Escherichia coli BL21 (DE3) pLysS cells
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expressed in Escherichia coli BL21(DE3) pLysS cells and in a Arabidopsis thaliana mutant (ugt78d2) deficient in anthocyanidin and flavonol 3-O-glucosyltransferase activity
expressed in Escherichia coli BL21(DE3)RIL cells
expressed in Escherichia coli DE3 pLys S cells
expressed in Escherichia coli DH5alpha cells
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expression in Escherichia coli
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), selenomethionine-labeled enzyme in Escherichia coli strain B834(DE3)
gene RF5, functional expression in Escherichia coli
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gene VvGT1, expression of His-tagged enzyme in Escherichia coli
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
enzyme expression is induced correspondingly by exogenous elicitors including gibberellic acid (about 2.5fold increased expression at 1 mg/l) and sucrose (about 5fold increased expression at 120 g/l)
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enzyme gene expression is detected only after the beginning of coloring stage or yellow stage, and is highest in the red pericarps
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ethylene better stimulates expression of the ufgt promoter in the dark than under light
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the enzyme is expressed not only at the sites of anthocyanin biosynthesis, but also at sites of flavonol glycoside biosynthesis
transcriptional activity is higher expressed in the red color of Phalaenopsis cultivars than in the white cultivars. In the red labellum of Phalaenopsis, the enzyme also shows higher expression levels than that in the white perianth
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treatment of grape berries with 1-methylcyclopropene inhibits ufgt mRNA accumulation
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while there is little enzyme expression before veraison, these transcripts begin to accumulate at week 8 and 10 after flowering to reach maximal levels at week 12 after flowering and decreases thereafter
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D121A
site-directed mutagenesis, inactive mutant
D121N
site-directed mutagenesis, inactive mutant
E381A
site-directed mutagenesis, inactive mutant
H22A
site-directed mutagenesis, inactive mutant
D374A
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site-directed mutagenesis, inactive mutant
H20A
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site-directed mutagenesis, inactive mutant
Q375H
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Q375N
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Q375N/T141A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T141A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme