BRENDA - Enzyme Database show
show all sequences of 1.14.14.83

Functional expression of a putative geraniol 8-hydroxylase by reconstitution of bacterially expressed plant CYP76F45 and NADPH-cytochrome P450 reductase CPR I from Croton stellatopilosus Ohba

Sintupachee, S.; Promden, W.; Ngamrojanavanich, N.; Sitthithaworn, W.; De-Eknamkul, W.; Phytochemistry 118, 204-215 (2015)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
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
Croton stellatopilosus
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
membrane
membrane-bound; membrane-bound
Croton stellatopilosus
16020
-
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
geraniol + [reduced NADPH-hemoprotein reductase] + O2
Croton stellatopilosus
-
(6E)-8-hydroxygeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
Croton stellatopilosus
-
18-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
Croton stellatopilosus Ohba
-
18-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Croton stellatopilosus
A0A0A7DLM4
-
-
Croton stellatopilosus
A0A0A8KYK4
cf. EC 1.6.2.4
-
Croton stellatopilosus Ohba
A0A0A7DLM4
-
-
Source Tissue
Source Tissue
Commentary
Organism
Textmining
leaf
;
Croton stellatopilosus
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
farnesol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus
12-hydroxyfarnesol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
farnesol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus Ohba
12-hydroxyfarnesol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
geraniol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus
(6E)-8-hydroxygeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
geraniol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus Ohba
(6E)-8-hydroxygeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus
18-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus Ohba
18-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus
16-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus Ohba
16-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
Subunits
Subunits
Commentary
Organism
?
x * 56700, about, sequence calculation; x * 56700, calculated, Cyp76F45; x * 78700, about, sequence calculation, x * 96000, recombinant His-tagged enzyme, SDS-PAGE
Croton stellatopilosus
Cofactor
Cofactor
Commentary
Organism
Structure
cytochrome P450
a cytochrome P450 monooxygenase; an NADPH-cytochrome P450 reductase
Croton stellatopilosus
FAD
-
Croton stellatopilosus
FMN
-
Croton stellatopilosus
heme
-
Croton stellatopilosus
NADPH
an NADPH-cytochrome P450 reductase
Croton stellatopilosus
pI Value
Organism
Commentary
pI Value Maximum
pI Value
Croton stellatopilosus
sequence calculation
-
5.27
Croton stellatopilosus
calculated, Cyp76F45
-
8.6
Croton stellatopilosus
sequence calculation
-
8.63
Cloned(Commentary) (protein specific)
Commentary
Organism
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
Croton stellatopilosus
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
Croton stellatopilosus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
cytochrome P450
an NADPH-cytochrome P450 reductase
Croton stellatopilosus
cytochrome P450
a cytochrome P450 monooxygenase
Croton stellatopilosus
FAD
-
Croton stellatopilosus
FMN
-
Croton stellatopilosus
heme
-
Croton stellatopilosus
NADPH
an NADPH-cytochrome P450 reductase
Croton stellatopilosus
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
membrane
membrane-bound
Croton stellatopilosus
16020
-
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
geraniol + [reduced NADPH-hemoprotein reductase] + O2
Croton stellatopilosus
-
(6E)-8-hydroxygeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
Croton stellatopilosus
-
18-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
Croton stellatopilosus Ohba
-
18-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
leaf
-
Croton stellatopilosus
-
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
farnesol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus
12-hydroxyfarnesol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
farnesol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus Ohba
12-hydroxyfarnesol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
geraniol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus
(6E)-8-hydroxygeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
geraniol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus Ohba
(6E)-8-hydroxygeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus
18-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus Ohba
18-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus
16-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
geranylgeraniol + [reduced NADPH-hemoprotein reductase] + O2
-
743431
Croton stellatopilosus Ohba
16-hydroxygeranylgeraniol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 78700, about, sequence calculation, x * 96000, recombinant His-tagged enzyme, SDS-PAGE
Croton stellatopilosus
?
x * 56700, about, sequence calculation; x * 56700, calculated, Cyp76F45
Croton stellatopilosus
pI Value (protein specific)
Organism
Commentary
pI Value Maximum
pI Value
Croton stellatopilosus
sequence calculation
-
5.27
Croton stellatopilosus
calculated, Cyp76F45
-
8.6
Croton stellatopilosus
sequence calculation
-
8.63
General Information
General Information
Commentary
Organism
evolution
enzyme CsCPR I belongs to Class I of dicotyledonous CPRs; enzyme CYP76F45 belongs to the CYP76 family of dicotyledonous CPR class I
Croton stellatopilosus
additional information
the enzyme contains specific oxygen- and heme-binding sites; the enzyme contains two FMN-, FAD-, and NADPH-binding sites and one P450-binding site
Croton stellatopilosus
physiological function
enzymes CYP97C27 and CPR I function as the enzyme geraniol-8-hydroxylase (G8H), which is likely to be involved in the biosynthesis of the indole alkaloid 19-E-vallesamine in Croton stellatopilosus; enzymes CYP97C27 and CPR I function as the enzyme geraniol-8-hydroxylase (G8H), which is likely to be involved in the biosynthesis of the indole alkaloid 19-E-vallesamine in Croton stellatopilosus
Croton stellatopilosus
General Information (protein specific)
General Information
Commentary
Organism
evolution
enzyme CsCPR I belongs to Class I of dicotyledonous CPRs
Croton stellatopilosus
evolution
enzyme CYP76F45 belongs to the CYP76 family of dicotyledonous CPR class I
Croton stellatopilosus
additional information
the enzyme contains two FMN-, FAD-, and NADPH-binding sites and one P450-binding site
Croton stellatopilosus
additional information
the enzyme contains specific oxygen- and heme-binding sites
Croton stellatopilosus
physiological function
enzymes CYP97C27 and CPR I function as the enzyme geraniol-8-hydroxylase (G8H), which is likely to be involved in the biosynthesis of the indole alkaloid 19-E-vallesamine in Croton stellatopilosus
Croton stellatopilosus
Other publictions for EC 1.14.14.83
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
746070
Balusamy
-
Overexpression of geraniol 10 ...
Panax ginseng
Plant Growth Regul.
81
305-316
2017
-
-
1
-
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-
-
-
1
-
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1
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1
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5
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1
1
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2
2
2
2
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743812
Pandey
Fungal endophytes of Catharan ...
Catharanthus roseus
Sci. Rep.
6
26583
2016
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1
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1
1
1
1
-
-
743431
Sintupachee
Functional expression of a pu ...
Croton stellatopilosus, Croton stellatopilosus Ohba
Phytochemistry
118
204-215
2015
-
-
1
-
-
-
-
-
1
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3
-
8
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1
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9
1
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5
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3
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2
6
-
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2
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3
-
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-
-
-
2
-
-
9
2
-
-
-
-
-
-
-
3
-
3
6
-
-
-
746441
Cui
Co-overexpression of geraniol ...
Catharanthus roseus
Sci. Rep.
5
8227
2015
-
3
1
-
1
-
-
-
-
-
-
1
-
3
-
-
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-
-
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-
1
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1
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3
1
1
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1
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1
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-
1
-
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-
-
2
2
-
-
-
743890
Liu
Enhancement of vindoline and ...
Catharanthus roseus
World J. Microbiol. Biotechnol.
30
175-180
2014
-
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
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-
2
-
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1
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2
-
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-
-
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-
-
-
-
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-
1
1
1
1
-
-
745686
Hoefer
Geraniol hydroxylase and hydr ...
Catharanthus roseus
Metab. Eng.
20
221-232
2013
-
-
1
1
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
2
-
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-
1
-
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1
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1
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-
-
-
-
-
2
-
-
-
-
1
-
-
-
-
-
1
1
-
-
-
718240
Huang
Differential expression and fu ...
Catharanthus roseus
Plant Sci.
190
16-23
2012
-
-
1
-
1
-
-
-
-
-
-
1
-
1
-
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1
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1
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1
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1
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1
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1
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1
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1
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1
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-
-
-
2
2
-
-
-
746239
Pan
Overexpression of ORCA3 and G ...
Catharanthus roseus
PLoS ONE
7
e43038
2012
-
-
-
-
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1
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1
1
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-
715234
Sung
Functional expression of geran ...
Catharanthus roseus
J. Agric. Food Chem.
59
4637-4643
2011
-
-
1
-
-
-
-
1
1
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1
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1
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1
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2
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1
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1
1
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1
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1
1
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1
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1
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2
-
1
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1
1
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-
1
1
-
1
1
716475
Poutrain
Molecular cloning and characte ...
Catharanthus roseus
Plant Biol.
13
36-41
2011
-
-
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1
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2
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2
2
2
2
-
-
714525
Wang
Cloning and functional analysi ...
Swertia mussotii
Biosci. Biotechnol. Biochem.
74
1583-1590
2010
-
-
1
-
-
-
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1
-
1
2
-
5
-
-
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-
-
3
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3
1
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1
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1
1
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1
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1
2
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-
-
3
-
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3
1
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-
-
-
-
-
-
-
1
3
3
1
-
-
716102
Peebles
The expression of 1-deoxy-d-xy ...
Catharanthus roseus
Metab. Eng.
13
234-240
2010
-
-
1
-
1
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-
-
-
-
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2
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1
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1
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1
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1
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-
-
-
-
2
2
-
-
-
716501
Wang
Overexpression of G10H and ORC ...
Catharanthus roseus
Plant Cell Rep.
29
887-894
2010
-
-
1
-
1
-
-
-
-
-
-
1
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1
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1
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1
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1
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1
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1
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-
1
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1
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-
-
-
-
-
-
-
2
2
2
2
-
-
716447
Oudin
-
The iridoid pathway in Cathara ...
Catharanthus roseus
Phytochem. Rev.
6
259-276
2007
-
-
1
-
-
-
-
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-
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2
-
1
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-
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2
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718213
Burlat
Co-expression of three MEP pat ...
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718237
Collu
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Catharanthus roseus
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162
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Collu
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Hallahan
Cytochrome P-450 in plant/inse ...
Catharanthus roseus, Nepeta racemosa
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1995
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Meijer
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Catharanthus roseus, no activity in Tabernaemontana pandacaqui, Tabernaemontana divaricata
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1993
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744287
Madyastha
Characterization of a cytochr ...
Catharanthus roseus
Biochemistry
15
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1976
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