Information on EC 1.3.1.77 - anthocyanidin reductase

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The expected taxonomic range for this enzyme is: Spermatophyta

EC NUMBER
COMMENTARY hide
1.3.1.77
-
RECOMMENDED NAME
GeneOntology No.
anthocyanidin reductase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
a flavan-3-ol + 2 NAD(P)+ = an anthocyanidin + 2 NAD(P)H + H+
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
redox reaction
-
reduction
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
2,3-cis-flavanols biosynthesis
-
-
Biosynthesis of secondary metabolites
-
-
Flavonoid biosynthesis
-
-
proanthocyanidins biosynthesis from flavanols
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-
SYSTEMATIC NAME
IUBMB Comments
flavan-3-ol:NAD(P)+ oxidoreductase
Forms 2,3-cis-flavan-3-ols. The isomeric 2,3-trans-flavan-3-ols are formed from flavan-3,4-diols by EC 1.17.1.3 leucoanthocyanidin reductase. While the enzyme from the legume Medicago truncatula (MtANR) uses both NADPH and NADH as reductant, that from the crucifer Arabidopsis thaliana (AtANR) uses only NADPH. Also, while the substrate preference of MtANR is cyanidin>pelargonidin>delphinidin, the reverse preference is found with AtANR.
CAS REGISTRY NUMBER
COMMENTARY hide
93389-48-1
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Arabidopsis thaliana Wassilevskija-2 ecotype
gene BANYULS orthologue, BAN
UniProt
Manually annotated by BRENDA team
gene BANYULS orthologue, four copies of BAN, two originating from each diploid progenitor
UniProt
Manually annotated by BRENDA team
gene BANYULS orthologue, four copies of BAN, two originating from each diploid progenitor
UniProt
Manually annotated by BRENDA team
gene BANYULS orthologue, BAN
D0QXJ3, D0QXJ3 and D0QXJ4
UniProt
Manually annotated by BRENDA team
gene BANYULS orthologue, BAN
UniProt
Manually annotated by BRENDA team
gene BANYULS orthologue, BAN
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
cultivar Hiratanenashi
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
brown-fiber cotton line T586
UniProt
Manually annotated by BRENDA team
cultivar Golden Delicious, Niedzwetz-kyana, Weirouge, Klarapfel, Red Delicious, Elstar, Holsteiner Cox, Pinova and Pilot
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Vitis vinifera Cabernet Sauvignon
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
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
(+)-epicatechin + 2 NADP+
cyanidin + 2 NADPH + 2 H+
show the reaction diagram
(-)-catechin + 2 NADP+
cyanidin + 2 NADPH + 2 H+
show the reaction diagram
(2S,3R)-flavan-3-ol + 2 NADP+
an anthocyanidin + 2 NADPH + H+
show the reaction diagram
-
Vitis vinifera anthocyanidin reductase, ANR, catalyzes an NADPH-dependent double reduction of anthocyanidins producing a mixture of (2S,3R)- and (2S,3S)-flavan-3-ols, product identification, overview. At pH 7.5 and 30C, the first hydride transfer to anthocyanidin is irreversible, and no intermediate is released during catalysis. ANR reverse activity is assessed in the presence of excess NADP+
-
-
r
(2S,3S)-flavan-3-ol + 2 NADP+
an anthocyanidin + 2 NADPH + H+
show the reaction diagram
-
Vitis vinifera anthocyanidin reductase, ANR, catalyzes an NADPH-dependent double reduction of anthocyanidins producing a mixture of (2S,3R)- and (2S,3S)-flavan-3-ols, product identification, overview. At pH 7.5 and 30C, the first hydride transfer to anthocyanidin is irreversible, and no intermediate is released during catalysis. ANR reverse activity is assessed in the presence of excess NADP+
-
-
r
2,3-cis-flavan-3-ol + NAD(P)+
anthocyanidin + NAD(P)H + H+
show the reaction diagram
the enzyme is involved in formation of condensed tannins. The enzyme competes with anthocyanidin synthase, for the pool of flavan-3,4-diol
-
-
?
anthocyanidin + NAD(P)H
2,3-cis-flavan-3-ol + NAD(P)+
show the reaction diagram
anthocyanidin + NAD(P)H + H+
2,3-cis-flavan-3-ol + NAD(P)+
show the reaction diagram
cyanidin + NAD(P)H + H+
(-)-epicatechin + NAD(P)+
show the reaction diagram
cyanidin + NADPH
(-)-epicatechin + NAD(P)+
show the reaction diagram
cyanidin + NADPH
epicatechin + NADP+
show the reaction diagram
-
-
-
-
?
cyanidin + NADPH + H+
(+)-epicatechin + NADP+
show the reaction diagram
enzyme produces a 50:50 mixture of 2S,3S-flavan-3-ol and 2S,3R-flavan-3-ol, i.e. (+)-epicatechin and (-)-catechin
-
-
?
cyanidin + NADPH + H+
(-)-catechin + NADP+
show the reaction diagram
enzyme produces a 50:50 mixture of 2S,3S-flavan-3-ol and 2S,3R-flavan-3-ol, i.e. (+)-epicatechin and (-)-catechin
-
-
?
cyanidin + NADPH + H+
(-)-epicatechin + NADP+
show the reaction diagram
cyanidin + NADPH + H+
(2S,3R)-3',4',5,7-tetrahydroxyflavan-3-ol + NADP+
show the reaction diagram
-
enzyme produces a 50:50 mixture of (2S,3S)-flavan-3-ol and (2S,3R)-flavan-3-ol, i.e. (+)-epicatechin and (-)-catechin
-
-
?
cyanidin + NADPH + H+
(2S,3S)-3',4',5,7-tetrahydroxyflavan-3-ol + NADP+
show the reaction diagram
-
enzyme produces a 50:50 mixture of (2S,3S)-flavan-3-ol and (2S,3R)-flavan-3-ol, i.e. (+)-epicatechin and (-)-catechin
-
-
?
cyanidin + NADPH + H+
epicatechin + NADP+
show the reaction diagram
-
-
-
?
delphinidin + 2 NADPH + H+
(-)-epigallocatechin + 2 NADP+
show the reaction diagram
delphinidin + NADPH
(-)-epigallocatechin + NAD(P)+
show the reaction diagram
-
changes in the concentration of products and coenzyme in the ANR assay are determined by thin layer chromatography (TLC), HPLC, mass spectrometry (MS) and UV spectrophotometry
-
-
?
delphinidin + NADPH
(-)-epigallocatechin + NADP+
show the reaction diagram
-
-
-
-
?
delphinidin + NADPH + H+
epigallocatechin + NADP+
show the reaction diagram
-
-
-
?
delphinidin + NADPH + H+
gallocatechin + NADP+
show the reaction diagram
-
-
-
?
NADPH + H+ + cyanidin
NADP+ + (-)-epicatechin
show the reaction diagram
-
-
-
-
?
NADPH + H+ + delphinidin
NADP+ + (-)-epigallocatechin
show the reaction diagram
NADPH + H+ + pelargonidin
NADP+ + (-)-epiafzelechin
show the reaction diagram
-
-
-
-
?
NADPH + H+ + petunidin
NADP+ + ?
show the reaction diagram
-
-
-
-
?
pelargonidin + 2 NADPH + H+
(-)-epiafzelechin + 2 NADP+
show the reaction diagram
pelargonidin + NAD(P)H + H+
epiafzelechin + NAD(P)+
show the reaction diagram
-
-
-
-
?
pelargonidin + NADPH + H+
afzelechin + NADP+
show the reaction diagram
-
-
-
?
pelargonidin + NADPH + H+
epiafzelechin + NADP+
show the reaction diagram
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
(+)-epicatechin + 2 NADP+
cyanidin + 2 NADPH + 2 H+
show the reaction diagram
(-)-catechin + 2 NADP+
cyanidin + 2 NADPH + 2 H+
show the reaction diagram
(2S,3R)-flavan-3-ol + 2 NADP+
an anthocyanidin + 2 NADPH + H+
show the reaction diagram
-
Vitis vinifera anthocyanidin reductase, ANR, catalyzes an NADPH-dependent double reduction of anthocyanidins producing a mixture of (2S,3R)- and (2S,3S)-flavan-3-ols, product identification, overview. At pH 7.5 and 30C, the first hydride transfer to anthocyanidin is irreversible, and no intermediate is released during catalysis. ANR reverse activity is assessed in the presence of excess NADP+
-
-
r
(2S,3S)-flavan-3-ol + 2 NADP+
an anthocyanidin + 2 NADPH + H+
show the reaction diagram
-
Vitis vinifera anthocyanidin reductase, ANR, catalyzes an NADPH-dependent double reduction of anthocyanidins producing a mixture of (2S,3R)- and (2S,3S)-flavan-3-ols, product identification, overview. At pH 7.5 and 30C, the first hydride transfer to anthocyanidin is irreversible, and no intermediate is released during catalysis. ANR reverse activity is assessed in the presence of excess NADP+
-
-
r
2,3-cis-flavan-3-ol + NAD(P)+
anthocyanidin + NAD(P)H + H+
show the reaction diagram
Q84XT1
the enzyme is involved in formation of condensed tannins. The enzyme competes with anthocyanidin synthase, for the pool of flavan-3,4-diol
-
-
?
anthocyanidin + NAD(P)H
2,3-cis-flavan-3-ol + NAD(P)+
show the reaction diagram
cyanidin + NADPH + H+
(+)-epicatechin + NADP+
show the reaction diagram
Q5FB34
enzyme produces a 50:50 mixture of 2S,3S-flavan-3-ol and 2S,3R-flavan-3-ol, i.e. (+)-epicatechin and (-)-catechin
-
-
?
cyanidin + NADPH + H+
(-)-catechin + NADP+
show the reaction diagram
Q5FB34
enzyme produces a 50:50 mixture of 2S,3S-flavan-3-ol and 2S,3R-flavan-3-ol, i.e. (+)-epicatechin and (-)-catechin
-
-
?
cyanidin + NADPH + H+
(2S,3R)-3',4',5,7-tetrahydroxyflavan-3-ol + NADP+
show the reaction diagram
-
enzyme produces a 50:50 mixture of (2S,3S)-flavan-3-ol and (2S,3R)-flavan-3-ol, i.e. (+)-epicatechin and (-)-catechin
-
-
?
cyanidin + NADPH + H+
(2S,3S)-3',4',5,7-tetrahydroxyflavan-3-ol + NADP+
show the reaction diagram
-
enzyme produces a 50:50 mixture of (2S,3S)-flavan-3-ol and (2S,3R)-flavan-3-ol, i.e. (+)-epicatechin and (-)-catechin
-
-
?
cyanidin + NADPH + H+
epicatechin + NADP+
show the reaction diagram
Q6DV46
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD(P)H
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(+)-catechin
-
0.5 mM, 50% inhibition
(+/-)-dihydroquercetin
-
0.025 mM
abscisic acid
-
decreases ANR activity and represses the expression a few days after application
Na+
-
above 200 mM
quercetin
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00282 - 0.0739
cyanidin
0.0142 - 0.0498
delphinidin
0.94
NADH
-
pH 6.0
0.097 - 0.45
NADPH
0.013 - 0.0528
pelargonidin
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0012 - 1.045
cyanidin
0.0026 - 1.34
delphinidin
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.111
NADPH
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
-
in 50 mM citrate/phosphate buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 8.9
-
pH 7.0: about 40% of maximal activity, pH 8.9: about 90% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
studies of binding at equilibrium
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
preferrently expressed
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36850
-
determined by SDS-PAGE and by MALDI mass spectrometry; theoretical mass without the initiator methionine
37000
-
calculated from cDNA
39000
-
SDS-PAGE, recombinant protein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
the structure of anthocyanidin reductase is described at a resolution of 2.2 A
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
extraction from plants, using a Sephadex G-25 column
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recombinant His6-tag fused GB1-ANR construct from Escherichia coli, Tev-mediated removal of the GB1 domain including the hexahistidine tag at its N-terminal end, and purification of Vv-ANR to homogeneity
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using a HiTrap Chelated HP column, the His-tag and GB1 domain are removed by digestion with TEV protease
-
using a HiTrap Chelated HP column, the His-tag is removed by digestion with TEV protease, the cleaved fraction is loaded on a gel filtration column
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
constitutive expression of the enzyme under control of the cauliflower mosaic virus 35S promoter in Nicotiana tabacum and Arabidopsis. Tobacco lines expressing the enzyme from Medicago trunculata lose the pink flower pigmentation characteristics of wild-type and empty vector control plants
expressed in Escherichia coli strain BL21
-
expressed in Escherichia coli. To investigate the function of PtrANR1, the open reading frame in sense or antisense orientation is introduced into Populus tomentosa Carr. plants for ectopic expression under the control of the cauliflower mosaic virus 35S promoter, respectively
-
expressed in Escherichia coli; expressed in Escherichia coli. Overexpressed in Nicotiana tabacum and Medicago truncatula for functional analysis
-
expressed in Escherichia coli; expressed in Escherichia coli; expressed in Escherichia coli; expressed in Escherichia coli; expressed in Escherichia coli; expressed in Escherichia coli; expressed in Escherichia coli
expressed in Malus domestica via Agrobacterium tumefaciens-mediated transformation
-
expression in Escherichia coli as a fusion protein with maltose-binding protein
for expression in Escherichia coli cells
gene BAN, DNA and amino acid sequence determination and analysis, sequence comparisons and genetic mapping, phylogenetic tree, overview
gene BAN, DNA and amino acid sequence determination and analysis, sequence comparisons and genetic mapping, phylogenetic tree, overview; gene BAN, DNA and amino acid sequence determination and analysis, sequence comparisons and genetic mapping, phylogenetic tree, overview
D0QXJ3 and D0QXJ4, D0QXJ3
gene BAN, DNA and amino acid sequence determination and analysis, sequence comparisons and genetic mapping, phylogenetic tree, overview; gene BAN, DNA and amino acid sequence determination and analysis, sequence comparisons and genetic mapping, phylogenetic tree, overview; gene BAN, DNA and amino acid sequence determination and analysis, sequence comparisons and genetic mapping, phylogenetic tree, overview; gene BAN, DNA and amino acid sequence determination and analysis, sequence comparisons and genetic mapping, phylogenetic tree, overview
genetic transformation of Arabidopsis thaliana with the Arabidopsis TT2 MYB transcription factor results in ectopic expression of the BANYULS gene, encoding anthocyanidin reductase, AHA10 encoding a P-type proton-pump and TT12 encoding a transporter involved in proanthocyanidin biosynthesis. When coupled with constitutive expression of PAP1, a positive regulator of anthocyanin biosynthesis, TT2 expression in Arabidopsis leads to the accumulation of proanthocyanidins, but only in a subset of cells in which the BANYULS promoter is naturally expressed. Ectopic expression of the maize Lc MYC transcription factor weakly induces AHA10 but does not induce BANYULS, TT12 or accumulation of proanthocyanidins
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into the vector pGEM-T and subsequently into pETGB_1a for expression in Escherichia coli BL21DE3 cells
production of a His6-tag fused GB1-ANR construct, expression of His6-tagged fused protein GB1-ANR in Escherichia coli
-
recombinantly expressed in Escherichia coli
-
the full-length VvANR cDNA is cloned into the vector pART7 and transformed into tobacco via the pART27 vector under the control of the CaMV 35S promoter
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
abscisic acid decreases ANR activity and represses the expression a few days after application
-
expression of anthocyanidin reductase is down-regulated in response to drought, abscisic acid and gibberellic acid
expression of anthocyanidin reductase is up-regulated in response to wounding
gene expression and enzymatic activity are higher in the developping leaves than in the mature leaves and lower than in the tender bud and first leaves
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
N185I/G214A
-
mutant shows minor reductions in activity. Calculated Km and Vmax values are not given as the enzymes are intensely inhibited by cyanidin concentrations above 0.1 mM
V122A/G214A
-
mutant shows minor reductions in activity. Calculated Km and Vmax values are not given as the enzymes are intensely inhibited by cyanidin concentrations above 0.1 mM
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
molecular biology
-
a method for the analysis of ANR activity using the detection of coenzyme is established