Information on EC 1.14.14.81 - flavanoid 3',5'-hydroxylase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.14.81
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RECOMMENDED NAME
GeneOntology No.
flavanoid 3',5'-hydroxylase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
a 3'-hydroxyflavanone + [reduced NADPH-hemoprotein reductase] + O2 = a 3',5'-dihydroxyflavanone + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
(1b)
-
-
-
a flavanone + 2 [reduced NADPH-hemoprotein reductase] + 2 O2 = a 3',5'-dihydroxyflavanone + 2 [oxidized NADPH-hemoprotein reductase] + 2 H2O
show the reaction diagram
overall reaction
-
-
-
a flavanone + [reduced NADPH-hemoprotein reductase] + O2 = a 3'-hydroxyflavanone + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
(1a)
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
flavonol biosynthesis
-
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Flavonoid biosynthesis
-
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Flavone and flavonol biosynthesis
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Biosynthesis of secondary metabolites
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SYSTEMATIC NAME
IUBMB Comments
flavanone,[reduced NADPH-hemoprotein reductase]:oxygen oxidoreductase (3',5'-dihydroxylating)
A heme-thiolate protein (P-450). The 3',5'-dihydroxyflavanone is formed via the 3'-hydroxyflavanone. In Petunia hybrida the enzyme acts on naringenin, eriodictyol, dihydroquercetin (taxifolin) and dihydrokaempferol (aromadendrin). The enzyme catalyses the hydroxylation of 5,7,4'-trihydroxyflavanone (naringenin) at either the 3' position to form eriodictyol or at both the 3' and 5' positions to form 5,7,3',4',5'-pentahydroxyflavanone (dihydrotricetin). The enzyme also catalyses the hydroxylation of 3,5,7,3',4'-pentahydroxyflavanone (taxifolin) at the 5' position, forming ampelopsin. NADH is not a good substitute for NADPH.
CAS REGISTRY NUMBER
COMMENTARY hide
94047-23-1
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
line '01'
SwissProt
Manually annotated by BRENDA team
var. pubilimba, crossings of cv. Yingshuan, from C. sinensis var. sinenesis, and cv. Beiyue Danzhu
UniProt
Manually annotated by BRENDA team
crossings of cv. Yingshuan and cv. Beiyue Danzhu from C. sinensis var. pubilimba
A0A1S5T865
UniProt
Manually annotated by BRENDA team
persimmon
UniProt
Manually annotated by BRENDA team
cv. Momokorin, pink flower plants
SwissProt
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
Met210 genetic variant with light purple flowers; w1 allele, Val210 genetic variant with white flowers
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-
Manually annotated by BRENDA team
Met210 genetic variant with light purple flowers
-
-
Manually annotated by BRENDA team
w1 allele, Val210 genetic variant with white flowers
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-
Manually annotated by BRENDA team
i.e. Gossypium maxicanum
SwissProt
Manually annotated by BRENDA team
blue potato bush
SwissProt
Manually annotated by BRENDA team
Nierembergia sp.
-
SwissProt
Manually annotated by BRENDA team
no activity in Chrysanthemum x morifolium
-
-
-
Manually annotated by BRENDA team
no activity in Rosa hybrida
-
-
-
Manually annotated by BRENDA team
cv. Blue Bicolor
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
cv. Big Chili
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene CYP75A31
UniProt
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
hybrid cultivar
Q9FS35
SwissProt
Manually annotated by BRENDA team
-
Q76LL4
SwissProt
Manually annotated by BRENDA team
Viola sp.
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2S)-naringenin + 2 + [reduced NADPH-hemoprotein reductase] + 2 O2
(2S)-5,7,3',4',5'-pentahydroxyflavanone + 2 + [oxidized NADPH-hemoprotein reductase] + 2 H2O
show the reaction diagram
2,3-(trans)-(+)-catechin + [reduced NADPH-hemoprotein reductase] + O2
2,3-(trans)-(+)-gallocatechin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
5,7,3',4'-tetrahydroxyflavanone + [reduced NADPH-hemoprotein reductase] + O2
5,7,3',4',5'-pentahydroxyflavanone + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
5,7,4'-trihydroxyflavanone + [reduced NADPH-hemoprotein reductase] + O2
5,7,3',4'-tetrahydroxyflavanone + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
5-deoxyleucopelargonidin + [reduced NADPH-hemoprotein reductase] + O2
? + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
?
apigenin + [reduced NADPH-hemoprotein reductase] + O2
5,7,3',4',5'-pentahydroxyflavone + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
apigenin + [reduced NADPH-hemoprotein reductase] + O2
?
show the reaction diagram
apigenin + [reduced NADPH-hemoprotein reductase] + O2
tricetin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
?
dihydrokaempferol + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
3',5'-dihydroxy-dihydrokaempferol + 2 [oxidized NADPH-hemoprotein reductase] + 2 H2O
show the reaction diagram
-
-
-
?
dihydrokaempferol + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
dihydromyricetin + 2 [oxidized NADPH-hemoprotein reductase] + 2 H2O
show the reaction diagram
dihydrokaempferol + [reduced NADPH-hemoprotein reductase] + O2
dihydromyricetin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
dihydrokaempferol + [reduced NADPH-hemoprotein reductase] + O2
dihydroquercetin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
dihydroquercetin + [reduced NADPH-hemoprotein reductase] + O2
5'-hydroxydihydroquercetin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
dihydroquercetin + [reduced NADPH-hemoprotein reductase] + O2
?
show the reaction diagram
-
-
-
-
?
dihydroquercetin + [reduced NADPH-hemoprotein reductase] + O2
dihydromyricetin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
kaempferol + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
5,7,3',4',5'-pentahydroxyflavone + 2 [oxidized NADPH-hemoprotein reductase] + 2 H2O
show the reaction diagram
kaempferol + [reduced NADPH-hemoprotein reductase] + O2
myricetin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
?
leucocyanidin + [reduced NADPH-hemoprotein reductase] + O2
leucodelphinidin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
naringenin + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
(2S)-5,7,3',4',5'-pentahydroxyflavanone + 2 + [oxidized NADPH-hemoprotein reductase] + 2 H2O
show the reaction diagram
naringenin + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
(2S)-5,7,3',4',5'-pentahydroxyflavanone + 2 [oxidized NADPH-hemoprotein reductase] + 2 H2O
show the reaction diagram
naringenin + [reduced NADPH-hemoprotein reductase] + O2
(2S)-eriodictyol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
naringenin + [reduced NADPH-hemoprotein reductase] + O2
pentahydroxyflavanone + [oxidized NADPH-hemoprotein reductase] + H2O
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
2,3-(trans)-(+)-catechin + [reduced NADPH-hemoprotein reductase] + O2
2,3-(trans)-(+)-gallocatechin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
apigenin + [reduced NADPH-hemoprotein reductase] + O2
?
show the reaction diagram
dihydrokaempferol + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
3',5'-dihydroxy-dihydrokaempferol + 2 [oxidized NADPH-hemoprotein reductase] + 2 H2O
show the reaction diagram
Q2PWV0, Q2PWV1, Q2PWV2, Q2PWV3, Q2PWV4
-
-
-
?
dihydrokaempferol + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
dihydromyricetin + 2 [oxidized NADPH-hemoprotein reductase] + 2 H2O
show the reaction diagram
dihydrokaempferol + [reduced NADPH-hemoprotein reductase] + O2
dihydromyricetin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
dihydrokaempferol + [reduced NADPH-hemoprotein reductase] + O2
dihydroquercetin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
dihydroquercetin + [reduced NADPH-hemoprotein reductase] + O2
5'-hydroxydihydroquercetin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
Q2PWV0, Q2PWV1, Q2PWV2, Q2PWV3, Q2PWV4
-
-
-
?
dihydroquercetin + [reduced NADPH-hemoprotein reductase] + O2
?
show the reaction diagram
-
-
-
-
?
dihydroquercetin + [reduced NADPH-hemoprotein reductase] + O2
dihydromyricetin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
leucocyanidin + [reduced NADPH-hemoprotein reductase] + O2
leucodelphinidin + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
-
-
-
-
?
naringenin + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
(2S)-5,7,3',4',5'-pentahydroxyflavanone + 2 + [oxidized NADPH-hemoprotein reductase] + 2 H2O
show the reaction diagram
naringenin + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
(2S)-5,7,3',4',5'-pentahydroxyflavanone + 2 [oxidized NADPH-hemoprotein reductase] + 2 H2O
show the reaction diagram
naringenin + [reduced NADPH-hemoprotein reductase] + O2
(2S)-eriodictyol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cytochrome b5
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cytochrome P450
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NADPH
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
a cytochrome P450 enzyme
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-aminobenzotriazole
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-
cytochrome c
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0.05 mM, complete inhibition
decyl-beta-D-glucopyranoside
-
dose-dependent inhibition
diethyldithiocarbamate
-
-
Digitonin
-
dose-dependent inhibition
dodecyl maltoside
-
dose-dependent inhibition
Emulgen 911
-
dose-dependent inhibition
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Emulgen 913
-
dose-dependent inhibition
heptyl-beta-D-glucopyranoside
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dose-dependent inhibition
hexyl-beta-D-glucopyranoside
-
dose-dependent inhibition
Lubrol Px
-
dose-dependent inhibition
Mega-10
-
dose-dependent inhibition
Mega-8
-
dose-dependent inhibition
Mega-9
-
dose-dependent inhibition
Nonidet P40
-
dose-dependent inhibition
nonyl-beta-D-glucopyranoside
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dose-dependent inhibition
octyl-beta-D-glucopyranoside
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dose-dependent inhibition
RNAi
reduces transcript levels, thus affecting flower color, acylated delphinidins (dominating in the wild-type) including gentiodelphin are decreased to 12.0% while non-acylated delphinidins are increased to 66.0 and 69.7% (15.2% in wild-type) of total anthocyanins in two transformed strains (clone 1 lilac, clone 15 pale blue)
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sodium cholate
-
dose-dependent inhibition
sodium deoxycholate
-
dose-dependent inhibition
Tetcyclacis
-
-
Triton X-100
-
dose-dependent inhibition
Triton X-114
-
dose-dependent inhibition
Tween 20
-
dose-dependent inhibition
Tween 80
-
dose-dependent inhibition
Zwittergent
-
dose-dependent inhibition
-
Zwittergent 3-08
-
dose-dependent inhibition
Zwittergent 3-10
-
dose-dependent inhibition
-
Zwittergent 3-12
-
dose-dependent inhibition
Zwittergent 3-14
-
dose-dependent inhibition
Zwittergent 3-16
-
dose-dependent inhibition
additional information
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a polyclonal antibody that inhibits higher plant NADPH-cytochrome p450 reductase inhibits the flavonoid 3',5'-hydroxylase. No effect by diethylpyrocarbonate or phenylmethylsulfonyl fluoride
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
maximal activation at 20 mM
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0058
apigenin
pH 7.0, 25°C
0.009
dihydrokaempferol
pH 7.0, 25°C
0.001
kaempferol
pH 7.0, 25°C
0.001
naringenin
pH 7.0, 25°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
13.4
apigenin
pH 7.0, 25°C
81.4
dihydrokaempferol
pH 7.0, 25°C
17
kaempferol
pH 7.0, 25°C
10.6
naringenin
pH 7.0, 25°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2300
apigenin
pH 7.0, 25°C
900
dihydrokaempferol
pH 7.0, 25°C
1700
kaempferol
pH 7.0, 25°C
10000
naringenin
pH 7.0, 25°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0000026
recombinant wild-type F3'5'H
additional information
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
high expression level in red petioles
Manually annotated by BRENDA team
F3'5'H activity in ray floret enzyme extracts of 3 developmental stages, overview
Manually annotated by BRENDA team
moderate expression level
Manually annotated by BRENDA team
Q3C210;
of young shoots
Manually annotated by BRENDA team
Q3C210;
young
Manually annotated by BRENDA team
-
expressed in tuber skin only in presence of the anthocyanin regulatory locus I
Manually annotated by BRENDA team
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
activity declines when microsomal suspensions are subjected to freezing and thawing
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, stable overnight in presence of 20% glycerol or sucrose at pH 7.5
-
75°C, storage in presence of 20% sucrose for periods of greater than 1 month results in significant loss of enzyme activity
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, genetic mapping, phylogenetic analysis, expression analysis and metabolic profiling, phenotypes; DNA and amino acid sequence determination and analysis, genetic mapping, phylogenetic analysis, expression analysis and metabolic profiling, phenotypes; DNA and amino acid sequence determination and analysis, genetic mapping, phylogenetic analysis, expression analysis and metabolic profiling, phenotypes; DNA and amino acid sequence determination and analysis, genetic mapping, phylogenetic analysis, expression analysis and metabolic profiling, phenotypes; DNA and amino acid sequence determination and analysis, genetic mapping, phylogenetic analysis, expression analysis and metabolic profiling, phenotypes
DNA and amino acid sequence determination and analysis, phylogenetic analysis of sequences of both enzymes indicate that F3',5'H is recruited from F3'H before the divergence of angiosperms and gymnosperms, overview
DNA and amino acid sequence determination and analysis, sequence comparison, genetic organization, and phylogenetic analysis
DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant enzyme overexpression in Petunia hybrida hybrid M1 x R27, which contains a mutated endogenous F3'5'H (HF1), no obvious phenotype
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expression in Dendrathemum grandiflora transgenic plants using the Agrobacterium tumefaciens stran ABA 4404 transfection method
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expression in Escherichia coli JM109
expression in Petunia hybrida; expression in Petunia hybrida
expression in Rosa hybrida transgenic plants, the mutant plants show the accumulation of a high percentage of delphinidin in selected cultuvars, while the wild-type lacks delphinidin and derivatives and the activity of F3'5'H
Viola sp.
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expression of wild-type enzyme and chimeric mutants in yeast strain INVSc 1 microsomes
functional expression of the enzyme as fusion enzyme with a P450 reductase leading to biosynthesis of plant-specific di- and trihydroxylated flavonols in Escherichia coli strain BL21(DE3), feeding experiments and determination of the flavonoid spectra in different recombinant bacterial lines in vivo, overview
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gene CYP75A31, DNA and amino acid sequence determination and analysis, functional expression in Saccharomyces cerevisiae microsomes using vector pYeDP60, real-time PCR expression analysis
gene F3',5'h, DNA and amino acid sequence determination and analysis, genomic structure, expression pattern analysis
Q3C210;
gene F3',5'H1, DNA and amino acid sequence determination and analysis, phylogenetic tree, developmental expression analysis, comparison of red and white cultivar enzyme expression levels, functional expression in Petunia hybrida altering the hosts' flower color and flavonoid composition
introduction of a flavonoid 3'5' hydroxylase sequence into Lotus root cultures. Expression of the transgene is associated with increased levels of condensed tannins, no alteration in polymer hydroxylation
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Met210 variant, DNA and amino acid sequence determination and analysis, semi-quantitative RT-PCR expression analysis; Val210 variant, DNA and amino acid sequence determination and analysis, semi-quantitative RT-PCR expression analysis
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one copy gene, DNA and amino acid sequence determination and analysis, sequence comparisons, genotyping, quantitative RT-PCR expression analysis
PCR-amplification of isolated cDNA
PCR-amplification, transformation into Agrobacterium tumefaciens EHA101 by electroporation used to transform the blue-flowered Gentiana triflora x Gentiana scabra cultivar Albireo via leaf disc infection
subcloning of AK14, encoding flavonoid-3',5'-hydroxylase, into a plant expression vector and transforming it to pink tobacco (Nicotiana tabacum cv. Petit Havana SR1) and pink petunia (var. Falcon), both of which originally lack the enzyme
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subcloning of TG1, encoding flavonoid-3',5'-hydroxylase, into a plant expression vector and transforming it to pink tobacco (Nicotiana tabacum cv. Petit Havana SR1) which originally lacks the enzyme
transcription profiling of different cultivars, overview
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when sense constructs are introduced into pink flower varieties that are deficient in the enzyme, transgenic plants show flower color changes from pink to magenta along with changes ihn anthocyanin composition. Some transgenic plants show novel pigmentation patterns, e.g. a star-shaped pattern. When sense constructs are introduced into blue flower petunia varieties, the flower color of the transgenic plants changes from deep blue to pale blue or even pale pink. Pigment composition analysis of transgenic plants suggests that the F3'5'H transgene not only creates or inhibits the biosynthetic pathway to 3',5'-hydroxylated anthocyanins but switches the pathway to 3',5'-hydroxylated or 3'-hydroxylated anthocyanins
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when VmFH1, encoding the flavonoid 3',5'-hydroxylase is expressed in transgenic petunia hybrida under the control of the cauliflower mosaic virus 35S promoter, some transgenic plants show drastic flower color alteration from red to deep red with deep purple sectors
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
berry skin accumulates cyanidin-based anthocyanins and delphinidin-based anthocyanins during development
-
chimeric RNAi technology silences transcription of enzyme, in clone 15 (pale blue) more than in clone 1 (lilac)
expression of gene CYP75A31 increases in response to nitrogen deprivation, in accordance with other genes in the phenylpropanoid pathway, overview
highest amount at candle stage, stage 3
in PCNA mutant, especially the relative expression compared to the substrate-competing flavonoid 3' hydroxylase sinks stronger with seasonal development than in the wild-type
transcript abundance of the flavonoid 3',5'-hydroxylase gene increases significantly during infection with fungus Endoconidiophora polonica
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F484Y/S487T
site-directed mutagenesis, altered substrate specificity compared to the wild-type enzyme
F48Y
site-directed mutagenesis, altered substrate specificity compared to the wild-type enzyme
S487A
site-directed mutagenesis, altered substrate specificity compared to the wild-type enzyme
S487Y
site-directed mutagenesis, altered substrate specificity compared to the wild-type enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
nutrition
additional information
understanding the regulation of flavonoid hydroxylases could be used to modify flavonoid composition of fruits
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