Information on EC 1.14.14.83 - geraniol 8-hydroxylase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.14.83
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RECOMMENDED NAME
GeneOntology No.
geraniol 8-hydroxylase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
geraniol + [reduced NADPH-hemoprotein reductase] + O2 = (6E)-8-hydroxygeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
secologanin and strictosidine biosynthesis
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Monoterpenoid biosynthesis
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Metabolic pathways
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Biosynthesis of secondary metabolites
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SYSTEMATIC NAME
IUBMB Comments
geraniol,[reduced NADPH-hemoprotein reductase]:oxygen oxidoreductase (8-hydroxylating)
Requires cytochrome P-450. Also hydroxylates nerol and citronellol, cf. EC 1.14.13.151 linalool 8-monooxygenase. The recommended numbering of geraniol gives 8-hydroxygeraniol as the product rather than 10-hydroxygeraniol as used by references 1-3. See prenol nomenclature {iupac/misc/prenol#p1::Pr-1}. The cloned enzyme also catalysed, but less efficiently, the 3'-hydroxylation of naringenin (cf. EC 1.14.13.21, flavonoid 3'-monooxygenase) [3].
CAS REGISTRY NUMBER
COMMENTARY hide
112198-82-0
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Manually annotated by BRENDA team
no activity in Tabernaemontana pandacaqui
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
; gene CYP76B10
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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
(R,S)-citronellol + [reduced NADPH-hemoprotein reductase] + O2
(6E)-8-hydroxycitronellol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
cis-nerol + [reduced NADPH-hemoprotein reductase] + O2
(6E)-8-hydroxynerol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
farnesol + [reduced NADPH-hemoprotein reductase] + O2
12-hydroxyfarnesol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
geraniol + [reduced NADPH-hemoprotein reductase] + O2
(6E)-8-hydroxygeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
geraniol + [reduced NADPH-hemoprotein reductase] + O2
10-hydroxygeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
16-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
18-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
nerol + [reduced NADPH-hemoprotein reductase] + O2
(6E)-8-hydroxynerol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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?
nerol + [reduced NADPH-hemoprotein reductase] + O2
10-hydroxynerol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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
(R,S)-citronellol + [reduced NADPH-hemoprotein reductase] + O2
(6E)-8-hydroxycitronellol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
cis-nerol + [reduced NADPH-hemoprotein reductase] + O2
(6E)-8-hydroxynerol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
geraniol + [reduced NADPH-hemoprotein reductase] + O2
(6E)-8-hydroxygeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
geraniol + [reduced NADPH-hemoprotein reductase] + O2
10-hydroxygeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
18-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
nerol + [reduced NADPH-hemoprotein reductase] + O2
(6E)-8-hydroxynerol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
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?
nerol + [reduced NADPH-hemoprotein reductase] + O2
10-hydroxynerol + [oxidized NADPH-hemoprotein reductase] + H2O
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cytochrome P450
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NADPH
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-chloromercuribenzoate
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0.1 mM, 17% residual activity
cantharanthine
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feedback inhibition
CO
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inhibition increases with increasing CO concentrations and is almost completely reversed with the aid of a light spurce between 420 and 450 nm
iodoacetamide
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10 mM, 87% residual activity
Metyrapone
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0.1 mM, 41% residual activity
N-ethylmaleimide
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1 mM, 58% residual activity
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
dithiothreitol
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3fold increase in activity in presence of dithiothreitol
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0055 - 0.0158
geraniol
0.011
nerol
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pH 7.2, 25C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.131 - 2.8
geraniol
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000008
geraniol
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pH 8.0, 30C, recombinant enzyme
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.66
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purified enzyme, pH 7.6, temperature not specified in the publication
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.7
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isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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moderate enzyme expression level
Manually annotated by BRENDA team
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parenchym
Manually annotated by BRENDA team
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actively growing cells around the root tip
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
55500
x * 55500, about, sequence calculation; x * 55500, recombinant G10H, SDS-PAGE
56000
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x * 56000, SDS-PAGE
56300
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x * 56300, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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secondary structure and three-dimensional structure analysis, overview
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
homology modeling. The geraniol molecule is positioned close to protoporphyrin with the double bond C6-C7 parallel to the heme plane
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native cholate-solubilized G10H about 8.3fold from suspension cell membranes by anion exchange and hydroxyapatite chromatography, and 2-aminooctyl agarose and a second step of hydrophobic interaction chromatography to homogeneity. Separation from the NADPH-cytochrome P-450 reductase, EC 1.6.2.4
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native enzyme about 12fold from membranes by anion exchange and omega-amino octyl agarose chromatography, followed by hydophobic interaction chromatography to homogeneity
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solubilization by treatment with sodium cholate, Renex-30, or Lubrol-WX
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning from a lambda-ZAPII library, recombinant expression in Saccharomyces cerevisiae strain YPH500 or in enzyme-deficient Catharathus roseus cell line MP183L, complementation
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co-expression of G10H with the NADPH-cytochrome P450 reductase from Arabidopsis thaliana in Spodoptera frugiperda Sf9 cell microsomes using the baculovirus transfection method, quantitative expression analysis
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expression in Saccharomyces cerevisiae
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expression in Spodoptera frugiperda SF9 cells, co-expression with NADPH:cytochrome P450 reductase NfCPR2 from Nothapodytes foetida and CPR2 from Arabidopsis thaliana
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gene CrG10H, recombinant expression in transgenic Ophiorrhiza pumila hairy roots using Agrobacterium rhizogenes strain C58C1 for transfection
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gene CsCPR I, DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis and tree, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3). Only simultaneous incubation of the membrane-bound enzymes CsCYP76F45 and CsCPR I with the substrate geraniol and the coenzyme NADPH allows 8-hydroxygeraniol formation; gene CYP76F45, DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis and tree, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3). Only simultaneous incubation of the membrane-bound enzymes CsCYP76F45 and CsCPR I with the substrate geraniol and the coenzyme NADPH allows 8-hydroxygeraniol formation
gene CYP76B10, DNA and amino acid sequence determination and analysis, semi-quantitative RT-PCR expression analysis, sequence comparison and phylogenetic tree, expression in Pichia pastoris and in Escherichia coli; SmG10H, DNA and amno acid sequence determination and analysis, sequence comparisons and phylogenetic tree, semiquantitative RT-PCR expression analysis, recombinant expression in Escherichia coli strain BL21 and in Pichia pastoris
gene G10H, DNA and amino acid sequence determination and analysis, phylogenetic analysis and tree, quantitative RT-PCR enzyme expression analysis. Recombinant overexpression of the ginseng geraniol 10-hydroxylase P450 gene in Arabidopsis thaliana ecotype CS60000, from vector pCAMBIA1390 under control of promoter CaMV 35S, causes terpenoid indole alkaloid dihydrositsirikine production and also confers enhanced resistance to Pseudomonas syringae pv. tomato DC3000
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gene g10h, quantitative real-time PCR expression analysis
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overexpression in Catharanthus roseus hairy roots via Agrobacterium rhizogenes 15834 transfection system, co-expression with the 1-deoxy-D-xylulose synthase, DXS, from Arabidopsis thaliana
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
alkaloid induction medium induces the enzyme, overview
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artemisinic acid is able to upregulate the transcriptions of geraniol 10-hydroxylase
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effect of differential T-DNA transfer on transcript accumulation of MIA biosynthetic pathway genes, expression of gene g10h is 15 to 130fold increased, overview
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expression is induced by fungal elicitor and methyl jasmonate
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G10H expression, recombinant in Escherichia coli, is upregulated by methyljasmonate 6-36 h after treatment; methyljasmonate induces CYP76B10 expression
G10H is repressed by ketonazole
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negative transcriptional regulation of geraniol 10-hydroxylase by calmodulin isozymes CAM1 and CAM2, overview
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the calmodulins CAM1 and CAM2 are required for monoterpene indole alkaloid biosynthesis in Catharanthus roseus cells by acting on regulation of expression of genes encoding enzymes that catalyse early steps of MIA biosynthesis, such as 1-deoxy-D-xylulose 5-phosphate reductoisomerase and geraniol 10-hydroxylase
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the expression of PgG10H is differentially induced in ginseng, not only during Pseudomonas syringae infection and wounding but also after exposure to methyl jasmonate and salt stress
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the stress hormone methyljasmonate strongly induces G10h gene expression coordinately with other terpenoid indole alkaloid biosynthesis genes in Catharanthus roseus cell culture
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the transcription level of gene g10h is increased 15 to 130fold by integration of T-DNA in host plant genome from root inducing Ri plasmid of Agrobacterium rhizogenes for induction of hariry root growth in second metabolite production, expression analysis in root clone CP68. Also the transcription of other genes involved in the monoterpene indole alkaloid biosynthetic pathway is altered, overview
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tissue-specific expression pattern and expression of PgG10H in response to various biotic and abiotic stresses
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upregulated in plants inoculated with endophytes
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various hormones and elicitors upregulate G10H and the monoterpene alkaloid biosynthesis pathways. G10H activity is induced by phenobarbital
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
reconstitution of the purified enzyme by addition of NADPH-cytochrome P-450 reductase, EC 1.6.2.4, and lipid extract
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis