BRENDA - Enzyme Database show
show all sequences of 1.1.1.318

The multiple phenylpropene synthases in both Clarkia breweri and Petunia hybrida represent two distinct protein lineages

Koeduka, T.; Louie, G.V.; Orlova, I.; Kish, C.M.; Ibdah, M.; Wilkerson, C.G.; Bowman, M.E.; Baiga, T.J.; Noel, J.P.; Dudareva, N.; Pichersky, E.; Plant J. 54, 362-374 (2008)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis
Petunia x hybrida
DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, expression of nontagged and of His-tagged CbEGS1 and CbEGS2 in Escherichia coli
Clarkia breweri
Engineering
Amino acid exchange
Commentary
Organism
F84V/I87Y
site-directed mutagenesis, the CbEGS1 mutant plants show increased isoeugenol production and an altered eugenol/isoeugenol ratio compared to the wild-type enzyme
Clarkia breweri
F86V/I89Y
site-directed mutagenesis, the CbEGS2 mutant plants show increased isoeugenol production and an altered eugenol/isoeugenol ratio compared to the wild-type enzyme
Clarkia breweri
Q86V/L89Y
site-directed mutagenesis, the mutant plants show increased isoeugenol production and an altered eugenol/isoeugenol ratio compared to the wild-type enzyme
Petunia x hybrida
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0933
-
coniferyl acetate
CbEGS1, pH 6.5, 28C
Clarkia breweri
0.2453
-
coniferyl acetate
PhEGS1, pH 6.5, 28C
Petunia x hybrida
0.3105
-
coniferyl acetate
CbEGS2, pH 6.5, 28C
Clarkia breweri
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
36000
-
x * 38000, recombinant nontagged CbEGS2, SDS-PAGE, x * 36000, recombinant nontagged CbEGS1, SDS-PAGE
Clarkia breweri
38000
-
x * 38000, recombinant nontagged CbEGS2, SDS-PAGE, x * 36000, recombinant nontagged CbEGS1, SDS-PAGE
Clarkia breweri
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
coniferyl acetate + NADPH + H+
Ocimum basilicum
-
eugenol + acetate + NADP+
-
-
?
coniferyl acetate + NADPH + H+
Clarkia breweri
-
eugenol + acetate + NADP+
-
-
?
coniferyl acetate + NADPH + H+
Petunia x hybrida
PhEGS1 converts coniferyl acetate to eugenol
eugenol + acetate + NADP+
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Clarkia breweri
-
-
-
Ocimum basilicum
-
-
-
Petunia x hybrida
-
cv. Mitchell
-
Purification (Commentary)
Commentary
Organism
native EGS 118fold from petals by alternating steps of anion exchange and hydrophobic interaction chromatography, co-purification with isoeugenol synthase, EC 1.1.1.319. Recombinant His-tagged CbEGS1 and CbEGS2 from Escherichia coli by nickel affinity chromatography
Clarkia breweri
Source Tissue
Source Tissue
Commentary
Organism
Textmining
flower
the highest specific activity levels of eugenol synthase are found in the stamens, followed closely by the pistil and petals
Clarkia breweri
-
flower
-
Ocimum basilicum
-
flower
PhEGS1 is expressed specifically in the scent-producing parts of the flowers, limbs and tube, but not in other parts of the flowers nor in leaves
Petunia x hybrida
-
petal
-
Clarkia breweri
-
pistil
-
Clarkia breweri
-
stamen
-
Clarkia breweri
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.98
-
purified native, wild-type CbEGS, pH 6.5, 28C
Clarkia breweri
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
coniferyl acetate + NADPH + H+
-
718214
Ocimum basilicum
eugenol + acetate + NADP+
-
-
-
?
coniferyl acetate + NADPH + H+
-
718214
Petunia x hybrida
eugenol + acetate + NADP+
-
-
-
?
coniferyl acetate + NADPH + H+
-
718214
Clarkia breweri
eugenol + acetate + NADP+
-
-
-
?
coniferyl acetate + NADPH + H+
PhEGS1 converts coniferyl acetate to eugenol
718214
Petunia x hybrida
eugenol + acetate + NADP+
-
-
-
?
coniferyl acetate + NADPH + H+
CbEGS1 is specific for isoeugenol production
718214
Clarkia breweri
eugenol + acetate + NADP+
-
-
-
?
coniferyl acetate + NADPH + H+
ObEGS1 uses a quinone methide intermediate-based mechanism to generate an intermediate to which the reductive transfer of the reducing hydride can then be easily accomplished. the enzyme acts on the substrate via a push-pull mechanism, removing the proton of the para hydroxyl group and promoting the cleavage of the acetyl group. In the resultant quinone-methide intermediate, the C7 atom serves as the acceptor of the hydride ion from NADPH
718214
Ocimum basilicum
eugenol + acetate + NADP+
-
-
-
?
Subunits
Subunits
Commentary
Organism
?
x * 38000, recombinant nontagged CbEGS2, SDS-PAGE, x * 36000, recombinant nontagged CbEGS1, SDS-PAGE
Clarkia breweri
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
28
-
assay at
Clarkia breweri
28
-
assay at
Petunia x hybrida
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.25
-
coniferyl acetate
CbEGS2, pH 6.5, 28C
Clarkia breweri
0.26
-
coniferyl acetate
CbEGS1, pH 6.5, 28C
Clarkia breweri
0.6
-
coniferyl acetate
PhEGS1, pH 6.5, 28C
Petunia x hybrida
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.5
-
assay at
Clarkia breweri
6.5
-
assay at
Petunia x hybrida
Cofactor
Cofactor
Commentary
Organism
Structure
NADPH
-
Clarkia breweri
NADPH
-
Petunia x hybrida
Cloned(Commentary) (protein specific)
Commentary
Organism
DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, expression of nontagged and of His-tagged CbEGS1 and CbEGS2 in Escherichia coli
Clarkia breweri
DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis
Petunia x hybrida
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADPH
-
Clarkia breweri
NADPH
-
Petunia x hybrida
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
F84V/I87Y
site-directed mutagenesis, the CbEGS1 mutant plants show increased isoeugenol production and an altered eugenol/isoeugenol ratio compared to the wild-type enzyme
Clarkia breweri
F86V/I89Y
site-directed mutagenesis, the CbEGS2 mutant plants show increased isoeugenol production and an altered eugenol/isoeugenol ratio compared to the wild-type enzyme
Clarkia breweri
Q86V/L89Y
site-directed mutagenesis, the mutant plants show increased isoeugenol production and an altered eugenol/isoeugenol ratio compared to the wild-type enzyme
Petunia x hybrida
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0933
-
coniferyl acetate
CbEGS1, pH 6.5, 28C
Clarkia breweri
0.2453
-
coniferyl acetate
PhEGS1, pH 6.5, 28C
Petunia x hybrida
0.3105
-
coniferyl acetate
CbEGS2, pH 6.5, 28C
Clarkia breweri
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
36000
-
x * 38000, recombinant nontagged CbEGS2, SDS-PAGE, x * 36000, recombinant nontagged CbEGS1, SDS-PAGE
Clarkia breweri
38000
-
x * 38000, recombinant nontagged CbEGS2, SDS-PAGE, x * 36000, recombinant nontagged CbEGS1, SDS-PAGE
Clarkia breweri
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
coniferyl acetate + NADPH + H+
Ocimum basilicum
-
eugenol + acetate + NADP+
-
-
?
coniferyl acetate + NADPH + H+
Clarkia breweri
-
eugenol + acetate + NADP+
-
-
?
coniferyl acetate + NADPH + H+
Petunia x hybrida
PhEGS1 converts coniferyl acetate to eugenol
eugenol + acetate + NADP+
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
native EGS 118fold from petals by alternating steps of anion exchange and hydrophobic interaction chromatography, co-purification with isoeugenol synthase, EC 1.1.1.319. Recombinant His-tagged CbEGS1 and CbEGS2 from Escherichia coli by nickel affinity chromatography
Clarkia breweri
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
flower
the highest specific activity levels of eugenol synthase are found in the stamens, followed closely by the pistil and petals
Clarkia breweri
-
flower
-
Ocimum basilicum
-
flower
PhEGS1 is expressed specifically in the scent-producing parts of the flowers, limbs and tube, but not in other parts of the flowers nor in leaves
Petunia x hybrida
-
petal
-
Clarkia breweri
-
pistil
-
Clarkia breweri
-
stamen
-
Clarkia breweri
-
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.98
-
purified native, wild-type CbEGS, pH 6.5, 28C
Clarkia breweri
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
coniferyl acetate + NADPH + H+
-
718214
Ocimum basilicum
eugenol + acetate + NADP+
-
-
-
?
coniferyl acetate + NADPH + H+
-
718214
Petunia x hybrida
eugenol + acetate + NADP+
-
-
-
?
coniferyl acetate + NADPH + H+
-
718214
Clarkia breweri
eugenol + acetate + NADP+
-
-
-
?
coniferyl acetate + NADPH + H+
PhEGS1 converts coniferyl acetate to eugenol
718214
Petunia x hybrida
eugenol + acetate + NADP+
-
-
-
?
coniferyl acetate + NADPH + H+
CbEGS1 is specific for isoeugenol production
718214
Clarkia breweri
eugenol + acetate + NADP+
-
-
-
?
coniferyl acetate + NADPH + H+
ObEGS1 uses a quinone methide intermediate-based mechanism to generate an intermediate to which the reductive transfer of the reducing hydride can then be easily accomplished. the enzyme acts on the substrate via a push-pull mechanism, removing the proton of the para hydroxyl group and promoting the cleavage of the acetyl group. In the resultant quinone-methide intermediate, the C7 atom serves as the acceptor of the hydride ion from NADPH
718214
Ocimum basilicum
eugenol + acetate + NADP+
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 38000, recombinant nontagged CbEGS2, SDS-PAGE, x * 36000, recombinant nontagged CbEGS1, SDS-PAGE
Clarkia breweri
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
28
-
assay at
Clarkia breweri
28
-
assay at
Petunia x hybrida
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.25
-
coniferyl acetate
CbEGS2, pH 6.5, 28C
Clarkia breweri
0.26
-
coniferyl acetate
CbEGS1, pH 6.5, 28C
Clarkia breweri
0.6
-
coniferyl acetate
PhEGS1, pH 6.5, 28C
Petunia x hybrida
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.5
-
assay at
Clarkia breweri
6.5
-
assay at
Petunia x hybrida
General Information
General Information
Commentary
Organism
physiological function
plants synthesize the volatile phenylpropene compounds eugenol and isoeugenol to serve in defense against herbivores and pathogens and to attract pollinators. Clarkia breweri flowers emit a mixture of eugenol and isoeugenol. Eugenol and isoeugenol differ in the position of the double bond in the propene side chain
Clarkia breweri
physiological function
plants synthesize the volatile phenylpropene compounds eugenol and isoeugenol to serve in defense against herbivores and pathogens and to attract pollinators. Petunia hybrida flowers emit mostly isoeugenol with small amounts of eugenol. Eugenol and isoeugenol differ in the position of the double bond in the propene side chain
Petunia x hybrida
General Information (protein specific)
General Information
Commentary
Organism
physiological function
plants synthesize the volatile phenylpropene compounds eugenol and isoeugenol to serve in defense against herbivores and pathogens and to attract pollinators. Clarkia breweri flowers emit a mixture of eugenol and isoeugenol. Eugenol and isoeugenol differ in the position of the double bond in the propene side chain
Clarkia breweri
physiological function
plants synthesize the volatile phenylpropene compounds eugenol and isoeugenol to serve in defense against herbivores and pathogens and to attract pollinators. Petunia hybrida flowers emit mostly isoeugenol with small amounts of eugenol. Eugenol and isoeugenol differ in the position of the double bond in the propene side chain
Petunia x hybrida
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.8
-
coniferyl acetate
CbEGS2, pH 6.5, 28C
Clarkia breweri
2.4
-
coniferyl acetate
PhEGS1, pH 6.5, 28C
Petunia x hybrida
2.8
-
coniferyl acetate
CbEGS1, pH 6.5, 28C
Clarkia breweri
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.8
-
coniferyl acetate
CbEGS2, pH 6.5, 28C
Clarkia breweri
2.4
-
coniferyl acetate
PhEGS1, pH 6.5, 28C
Petunia x hybrida
2.8
-
coniferyl acetate
CbEGS1, pH 6.5, 28C
Clarkia breweri
Other publictions for EC 1.1.1.318
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)
740112
Anand
Comparative functional charact ...
Ocimum basilicum, Ocimum gratissimum, Ocimum kilimandscharicum, Ocimum tenuiflorum
Biochim. Biophys. Acta
1864
1539-1547
2016
-
-
-
4
-
-
-
7
-
-
-
-
-
6
-
-
-
-
-
8
-
-
11
-
-
-
-
7
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
7
-
-
-
-
-
-
-
-
-
8
-
-
11
-
-
-
-
7
-
-
-
-
-
-
-
-
7
7
741204
Medina-Puche
An R2R3-MYB transcription fact ...
Fragaria x ananassa
Plant Physiol.
168
598-614
2015
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
740496
Gupta
Eugenol synthase genes in flor ...
Gymnadenia odoratissima, Gymnadenia x densiflora
Funct. Integr. Genomics
14
779-788
2014
-
-
-
-
-
-
-
-
-
-
2
-
-
10
-
-
-
-
-
4
-
-
8
2
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
8
-
-
8
4
-
-
-
-
-
-
-
4
-
-
-
-
-
-
739985
Koeduka
Enhancement of production of e ...
Petunia x hybrida
Biochem. Biophys. Res. Commun.
436
73-78
2013
-
-
-
-
-
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
718078
Hoffmann
Metabolic engineering in straw ...
Ocimum basilicum
Metab. Eng.
13
527-531
2011
-
-
1
-
1
-
-
-
-
-
-
3
-
1
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
1
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
718216
Koeduka
The lack of floral synthesis a ...
Petunia axillaris subsp. parodii
Plant J.
58
961-969
2009
-
-
1
-
-
-
-
-
-
-
-
1
-
10
-
-
-
-
-
1
-
-
2
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
718214
Koeduka
The multiple phenylpropene syn ...
Clarkia breweri, Ocimum basilicum, Petunia x hybrida
Plant J.
54
362-374
2008
-
-
2
-
3
-
-
3
-
-
2
3
-
8
-
-
1
-
-
6
1
-
6
1
2
-
-
3
2
-
-
2
-
-
-
-
-
2
2
-
3
-
-
-
-
3
-
-
2
3
-
-
-
1
-
6
1
-
6
1
2
-
-
3
2
-
-
-
-
2
2
-
3
3
717154
Vassao
A pinoresinol-lariciresinol re ...
Larrea tridentata
Arch. Biochem. Biophys.
465
209-218
2007
-
-
1
-
-
-
-
2
-
-
-
2
-
1
-
-
1
1
-
1
-
-
4
-
1
-
-
2
1
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
2
-
-
-
2
-
-
-
1
-
1
-
-
4
-
1
-
-
2
1
-
-
-
-
1
1
-
2
2
718254
Louie
Structure and reaction mechani ...
Ocimum basilicum
PLoS ONE
2
e993
2007
-
-
1
1
-
-
1
-
-
-
-
1
-
3
-
-
-
1
-
-
-
-
3
2
1
-
-
-
1
-
-
1
-
-
-
-
-
1
1
1
-
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
3
2
1
-
-
-
1
-
-
-
-
4
4
-
-
-
718314
Koeduka
Eugenol and isoeugenol, charac ...
Ocimum basilicum
Proc. Natl. Acad. Sci. USA
103
10128-10133
2006
-
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
-
1
-
1
-
-
2
-
1
-
-
-
1
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
-
2
-
1
-
-
-
1
-
-
-
-
1
1
-
-
-