Information on EC 2.4.1.115 - anthocyanidin 3-O-glucosyltransferase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.4.1.115
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RECOMMENDED NAME
GeneOntology No.
anthocyanidin 3-O-glucosyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
UDP-D-glucose + an anthocyanidin = UDP + an anthocyanidin-3-O-beta-D-glucoside
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexosyl group transfer
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
anthocyanidin modification (Arabidopsis)
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anthocyanin biosynthesis
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Anthocyanin biosynthesis
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anthocyanin biosynthesis (delphinidin 3-O-glucoside)
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anthocyanin biosynthesis (pelargonidin 3-O-glucoside)
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Biosynthesis of secondary metabolites
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Metabolic pathways
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rose anthocyanin biosynthesis II (via cyanidin 3-O-beta-D-glucoside)
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superpathway of anthocyanin biosynthesis (from cyanidin and cyanidin 3-O-glucoside)
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SYSTEMATIC NAME
IUBMB Comments
UDP-D-glucose:anthocyanidin 3-O-beta-D-glucosyltransferase
The anthocyanidin compounds cyanidin, delphinidin, peonidin and to a lesser extent pelargonidin can act as substrates. The enzyme does not catalyse glucosylation of the 5-position of cyanidin and does not act on flavanols such as quercetin and kaempferol (cf. EC 2.4.1.91 flavonol 3-O-glucosyltransferase). In conjunction with EC 1.14.11.19, leucocyanidin oxygenase, it is involved in the conversion of leucoanthocyanidin into anthocyanidin 3-glucoside. It may act on the pseudobase precursor of the anthocyanidin rather than on the anthocyanidin itself [3].
CAS REGISTRY NUMBER
COMMENTARY hide
65607-32-1
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Manually annotated by BRENDA team
gene Ct3GT-A
UniProt
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
cultivar Clark line
UniProt
Manually annotated by BRENDA team
cv. Dutch Red Hybrid
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Manually annotated by BRENDA team
Hort. cv. Blue Diamond
SwissProt
Manually annotated by BRENDA team
cultivars with non-red, pale red and red apple skin
UniProt
Manually annotated by BRENDA team
Chinese bayberry, white cultivar Shuijing (SJ), red cultivar Dongkui (DK), dark red-purple cultivar Biqi (BQ)
UniProt
Manually annotated by BRENDA team
enzyme has activity of both EC 2.4.1.115 and EC 2.4.1.116
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
tulip, var. Most Miles
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Manually annotated by BRENDA team
cultivar Concord
UniProt
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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conparison of structure and substrate specificity of UDP-glucose:anthocyanidin 3-O-glucosyltransferase, Ct3GT-A, from Clitoria ternatea and flavonoid glycosyltransferase, VvGT1, from Vitis vinifera detailed overview
malfunction
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two wild-type plants and two independent mutants of Medicago truncatula with altered anthocyanin content in leaves are compared at metabolite profile, gene structure and enzyme transcript levels. Flavonoid profiles show conserved levels of dihydroflavonols, leucoanthocyanidins and flavonols, while anthocyanidin, anthocyanin and isoflavone levels are lower in the mutants (up to 90% less) compared with the wild-types, phenotypes, overview. The enzyme is downregulated in the mutants compared to wild-type
metabolism
additional information
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the structure of Ct3GT-A shows a common folding topology, the GT-B fold, comprised of two Rossmann-like beta/alpha/beta domains and a cleft located between the N- and C-domains containing two cavities that are used as binding sites for the donor (UDP-Glc) and acceptor substrates, structure-function relationship analysis, overview. The donor-binding site conserved as a UGT signature PSPG motif is located in the C-domain of Ct3GT-A, and the C-terminal helix comprising residues 431-445 participates in forming the N-domain after crossing the cleft. The acceptor-binding site is formed mostly by the residues from the N-domain. Besides the hydrophobic residues Phe12, Phe116, Trp135, Tyr145, Phe192 and Leu196, the hydrophilic residues Asn137, Asp181 and Asp367 are arranged to form the acceptor-binding site
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
anthocyanidin + UDP-D-glucose
anthocyanidin 5-O-glucoside + UDP
show the reaction diagram
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-
-
-
?
anthocyanidin 5-O-glucoside + UDP-D-glucose
anthocyanidin-3,5-O-diglucoside + UDP
show the reaction diagram
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-
-
-
?
UDP-alpha-D-glucose + 3,7-dihydroxyflavone
UDP + 3-[(beta-D-glucopyranosyl)oxy]-7-hydroxyflavone
show the reaction diagram
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about 10% of the activity with cyanidin
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-
?
UDP-alpha-D-glucose + cyanidin
UDP + ?
show the reaction diagram
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
-
about 25% of the activity with cyanidin
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-
?
UDP-alpha-D-glucose + kaempferol
UDP + kaempferol 3-O-beta-D-glucoside
show the reaction diagram
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67% of the activity with cyanidin
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?
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-D-glucose + anthocyanidin
UDP + anthocyanidin-3-O-beta-D-glucoside
show the reaction diagram
-
-
-
?
UDP-D-glucose + cyanidin
UDP + cyanidin-3-O-beta-D-glucoside
show the reaction diagram
-
17% of the activity with delphinidin
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-
?
UDP-D-glucose + delphinidin
UDP + delphinidin-3-O-beta-D-glucoside
show the reaction diagram
UDP-D-glucose + malvidin
UDP + malvidin-3-O-beta-D-glucoside
show the reaction diagram
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87% of the activity with delphinidin
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-
?
UDP-D-glucose + pelargonidin
UDP + pelargonidin-3-O-beta-D-glucoside
show the reaction diagram
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29% of the activity with delphinidin
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-
?
UDP-D-glucose + peonidin
UDP + peonidin-3-O-beta-D-glucoside
show the reaction diagram
-
77% of the activity with delphinidin
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-
?
UDP-D-glucose + petunidin
UDP + petunidin-3-O-beta-D-glucoside
show the reaction diagram
UDP-glucose + (2R,3R)-dihydroquercetin
UDP + (2R,3R)-dihydroquercetin 3-O-beta-D-glucoside
show the reaction diagram
worst substrate
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-
?
UDP-glucose + anthocyanidin
UDP + anthocyanidin-3-O-beta-D-glucoside
show the reaction diagram
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-
-
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?
UDP-glucose + cyanidin
UDP + cyanidin-3-O-beta-D-glucoside
show the reaction diagram
UDP-glucose + delphinidin
UDP + delphinidin 3-O-beta-D-glucoside
show the reaction diagram
UDP-glucose + delphinidin
UDP + delphinidin 3-O-glucoside
show the reaction diagram
UDP-glucose + delphinidin
UDP + delphinidin-3-O-beta-D-glucoside
show the reaction diagram
UDP-glucose + fisetin
UDP + fisetin-3-O-beta-D-glucoside
show the reaction diagram
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16% of the activity with cyanidin
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?
UDP-glucose + isorhamnetin
UDP + isorhamnetin-3-O-beta-D-glucoside
show the reaction diagram
UDP-glucose + kaempferol
UDP + kaempferol-3-O-beta-D-glucoside
show the reaction diagram
UDP-glucose + malvidin
UDP + malvidin 3-O-beta-D-glucoside
show the reaction diagram
UDP-glucose + malvidin
UDP + malvidin 3-O-glucoside
show the reaction diagram
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-
-
?
UDP-glucose + malvidin
UDP + malvidin-3-O-beta-D-glucoside
show the reaction diagram
UDP-glucose + malvidin
UDP + quercetin-3-O-beta-D-glucoside
show the reaction diagram
best substrate
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-
?
UDP-glucose + myricetin
UDP + myricetin-3-O-beta-D-glucoside
show the reaction diagram
UDP-glucose + pelargonidin
UDP + pelargonidin 3-O-glucoside
show the reaction diagram
UDP-glucose + pelargonidin
UDP + pelargonidin-3-O-beta-D-glucoside
show the reaction diagram
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?
UDP-glucose + peonidin
UDP + peonidin 3-O-beta-D-glucoside
show the reaction diagram
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52% of the activity with cyanidin
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?
UDP-glucose + peonidin
UDP + peonidin-3-O-beta-D-glucoside
show the reaction diagram
UDP-glucose + petunidin
UDP + petunidin-3-O-beta-D-glucoside
show the reaction diagram
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-
-
?
UDP-glucose + quercetin
UDP + quercetin-3-O-beta-D-glucoside
show the reaction diagram
UDPalpha-D-glucose + galangin
UDP + galangin 3-O-bbeta-D-glucoside
show the reaction diagram
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about 15% of the activity with cyanidin
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?
UDPglucose + anthocyanidin
UDP + anthocyanidin-3-O-glucoside
show the reaction diagram
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involved in anthocyanine biosynthesis
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?
UDPglucose + cyanidin
UDP + cyanidin 3-O-beta-D-glucoside
show the reaction diagram
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?
UDPglucose + cyanidin
UDP + cyanidin 3-O-glucoside
show the reaction diagram
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best substrate
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?
UDPglucose + fisetin
UDP + fisetin 3-O-glucoside
show the reaction diagram
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about 10% of the activity with cyanidin
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?
UDPglucose + kaempferol
UDP + kaempferol 3-O-glucoside
show the reaction diagram
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about 5% of the activity with cyanidin
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?
UDPglucose + malvidin
UDP + malvidin 3-O-glucoside
show the reaction diagram
UDPglucose + myricetin
UDP + 3-O-glucoside myricetin
show the reaction diagram
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about 15% of the activity with cyanidin
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?
UDPglucose + peonidin
UDP + peonidin 3-O-beta-D-glucoside
show the reaction diagram
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about 85% of the activity with cyanidin
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?
UDPglucose + petunidin
UDP + petunidin 3-O-glucoside
show the reaction diagram
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about 35% of the activity with cyanidin
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?
UDPglucose + quercetin
UDP + quercetin 3-O-glucoside
show the reaction diagram
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about 15% of the activity with cyanidin
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?
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
UDP-alpha-D-glucose + cyanidin
UDP + cyanidin 3-O-beta-D-glucoside
show the reaction diagram
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first step of a anthocyanin branch
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?
UDP-D-glucose + anthocyanidin
UDP + anthocyanidin-3-O-beta-D-glucoside
show the reaction diagram
C9K224
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-
-
?
UDP-D-glucose + delphinidin
UDP + delphinidin-3-O-beta-D-glucoside
show the reaction diagram
UDP-D-glucose + petunidin
UDP + petunidin-3-O-beta-D-glucoside
show the reaction diagram
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-
-
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?
UDP-glucose + delphinidin
UDP + delphinidin 3-O-beta-D-glucoside
show the reaction diagram
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first step of a anthocyanin branch
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?
UDPglucose + anthocyanidin
UDP + anthocyanidin-3-O-glucoside
show the reaction diagram
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involved in anthocyanine biosynthesis
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?
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
no cofactors required
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
CaCl2
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2 mM, 18% inhibition
Cu2+
0.01 mM, over 80% reduction in quercetin glucosylation activity
cyanidin
cyanidin 3-O-beta-D-glucoside
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5 mM, product inhibition
HgCl2
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0.1 mM, 85% inhibition
NEM
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1.25 mM, 30% inhibition
UDP-glucose
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ZnCl2
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2 mM, 22% inhibition
additional information
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most of the activity is lost after ultrafiltration or (NH4)2SO4 precipitation, but it is restored or even enhanced by the addition of the ultrafiltrate or ascorbic acid to the assay. Other reducing agents are not able to replace ascorbic acid
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
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detergent required for maximal activity
2-Methoxyethanol
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activation
Cetrimide
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activation
cysteine
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detergent required for maximal activity
MrMYB1 transcription factor
strong correlation with anthocyanin content in ripe fruit, which is positively correlated with expression of the enzyme
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Triton X-100
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activation
Tween 20
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detergent required for maximal activity
Tween 80
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detergent required for maximal activity
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00479 - 0.4
cyanidin
0.016 - 0.13
delphinidin
0.174
kaempferol
recombinant enzyme, at pH 8.0 and 30°C
0.0357
malvidin
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0.13
pelargonidin
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0.00216 - 0.015
quercetin
0.2 - 1.9
UDP-glucose
0.41
UDPglucose
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000704
cyanidin
Vitis labrusca
A6N928
in 100 mM Tris-HCl pH 8.0, at 30°C
1.25
kaempferol
Glycine max
D3Y5N8
recombinant enzyme, at pH 8.0 and 30°C
0.000113
quercetin
Vitis labrusca
A6N928
in 100 mM Tris-HCl pH 8.0, at 30°C
0.000247
UDP-glucose
Vitis labrusca
A6N928
in 100 mM Tris-HCl pH 8.0, at 30°C
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 30°C
1072
0.052
quercetin
Vitis labrusca
A6N928
in 100 mM Tris-HCl pH 8.0, at 30°C
137
0.00027
UDP-glucose
Vitis labrusca
A6N928
in 100 mM Tris-HCl pH 8.0, at 30°C
64
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.035
cyanidin
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1
UDP-glucose
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0252
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8.8
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pH 6.0: about 44% of maximal activity, pH 8.8: about 60% of maximal activity, reaction with cyanidin and UDP-glucose
6.6 - 8
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50% of maximal activity at pH 6.6 and pH 8.0
6.6 - 9.4
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pH 6.6: about 40% of maximal activity, pH 9.4: about 50% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.54
calculated from sequence
5.6
-
calculated from amino acid sequence
6
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
pre-veraison exocarp
Manually annotated by BRENDA team
additional information
barely expressed in root and leaf, not in stem
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26000
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gel filtration
48043
x * 48043, calculated from amino acid sequence
49240
x * 49240, calculated from amino acid sequence
49560
calculated from sequence
49800
x * 49800, SDS-PAGE
49900
-
x * 49900, calculated from amino acid sequence
50000
x * 50000, SDS-PAGE
50100
x * 50100, calculated from amino acid sequence
56000
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gel filtration
60000
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1 * 60000, can exist as monomer or dimer, SDS-PAGE; 2 * 60000, can exist as monomer or dimer, SDS-PAGE
78000
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gel filtration
125000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
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2 * 60000, can exist as monomer or dimer, SDS-PAGE
monomer
-
1 * 60000, can exist as monomer or dimer, SDS-PAGE
additional information
-
the structure of Ct3GT-A shows a common folding topology, the GT-B fold, comprised of two Rossmann-like beta/alpha/beta domains and a cleft located between the N- and C-domains containing two cavities that are used as binding sites for the donor (UDP-Glc) and acceptor substrate, structure-function relationship analysis, overview
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified enzyme alone and in complex with anthocyanidin delphinidin and petunidin, and flavonol kaempferol, X-ray diffraction structure determination and analysis at 1.85 A, 2.55 A, 2.70 A, and 1.75 A resolution, respectively
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purified recombinant detagged enzyme, hanging drop vapor diffusion method, mixing of protein in 20 mM Tris-HCl, pH 7.4, 200 mM NaCl and 2 mM CaCl2, with reservoir solution containing 0.1 M sodium citrate tribasic dihydrate, pH 5.6, 0.2 M ammonium acetate and 26% w/v PEG 4000, equilibration against reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 1.85 A resolution
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
at least three freeze-thaw cycles can be tolerated without apparent loss of activity
repeated freeze-thawing inactivates
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-17°C, about 40% loss of activity within 3 months
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-20°C, recombinant enzyme is stable for at least 120 h
-80°C, recombinant enzyme is stable for many months
4°C or -20°C, 50% glycerol solution, 24 h, no significant loss of activity
4°C, about 40% loss of activity within 1 week
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4°C, recombinant enzyme is stable for at least 120 h
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
glutathione Sepharose column chromatography
Ni Sepharose column chromatography
partial
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recombinant
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recombinant His-tagged enzyme from Escherichia coli strain XL1 Blue by nickel affinity chromatography, removal of the N-terminal His-tag by digestion using recombinant enterokinase, followed by cation exchange chromatography, to homogeneity
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Talon metal affinity column chromatography, gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, sequence comparisons and genotyping of genes encoding enzymes of the anthocyanin metabolism
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expressed in Escherichia coli DE3 pLysS cells
expressed in Escherichia coli strain BL21 (DE3) cells and in Petunia grandiflora by Agrobacterium-mediated transformation
expressed in the Arabidopsis thaliana T-DNA mutant (ugt78d2) deficient in anthocyanidin and flavonol 3-O-beta-D-glucosyltransferase activity and in Escherichia coli BL21(DE3) pLysS cells
expression in Escherichia coli
gene Ct3GT-A, recombinant His-tagged ezyme expression in Escherichia coli strain XL1 Blue
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MrMYB1 transcription factor
optimized expression in Escherichia coli
the full-length open reading frame of FaGT1 is cloned into the expression vector pET-29a(1) for heterologous protein expression in Escherichia coli
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
17°C + UV-B radiation, 17°C without UV-B radiation, 27°C + UV-B radiation, 27°C without UV-B radiation
fruit bagging inhibits anthocyanin biosynthetic genes and expression of transcription factor MrMYB1, and subsequently anthocyanin accumulation, less colored cultivars contain a nonsense mutation in transcription factor MYB1
gene expression is highest 12 weeks after flowering in the exocarp
highest transcript level is found at 4.0 mg/l gibberelic acid treatment
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the more anthocyanin, the more strongly the enzyme is expressed, only expression in ripe fruit tissues, overexpression of transcription factor MrMYB1 stimulates anthocyanin accumulation
transcript level in the unpigmented buds is low, but increases during the pigment-accumulation stages and peaks in fully opened petals. Enzyme expression level is identical in red and pink petals and is lower in yellow petals, and rarely detectable in white petals
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
two wild-type plants and two independent mutants of Medicago truncatula with altered anthocyanin content in leaves are compared at metabolite profile, gene structure and enzyme transcript levels. Flavonoid profiles show conserved levels of dihydroflavonols, leucoanthocyanidins and flavonols, while anthocyanidin, anthocyanin and isoflavone levels are lower in the mutants (up to 90% less) compared with the wild-types, phenotypes, overview. The enzyme is downregulated in the mutants compared to wild-type
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
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expression of enzyme in Escherichia coli, together with dihydroflavonol 4-reductase, anthocyanidin synthase, flavanone 3beta-hydroxylase. production of pelargonidin 3-O-glucoside or cyanidin 3-O-glucoside from naringenin or eriodyctol, resp., via this recombinant plant pathway. Yields of 0.0056 mg/l pelargonidin 3-O-glucoside, or of 0.006 mg/l cyanidin 3-O-glucoside