Information on EC 4.2.3.108 - 1,8-cineole synthase

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The expected taxonomic range for this enzyme is: Gunneridae

EC NUMBER
COMMENTARY
4.2.3.108
-
RECOMMENDED NAME
GeneOntology No.
1,8-cineole synthase
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
geranyl diphosphate + H2O = 1,8-cineole + diphosphate
show the reaction diagram
-
-
-
-
geranyl diphosphate + H2O = 1,8-cineole + diphosphate
show the reaction diagram
substrate in left-hynded screw-sense orientation isomerizes to (3R)-linalyl diphosphate which in anti,endo-conformation cyclizes to the (4R)-alpha-terpinyl intermediate and thence to cineole following the addition of water
-
geranyl diphosphate + H2O = 1,8-cineole + diphosphate
show the reaction diagram
syn-stereochemistry for both C-O-bond-forming steps. Reaction starts with allylic rearrangement of geranyl diphosphate to (R)-linalyl diphosphate, followed by SN' cyclization to alpha-terpineol and proton-induced cyclization of the C=C-double bond and the tertiary hydroxyl groups
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
Metabolic pathways
-
monoterpene biosynthesis
-
Monoterpenoid biosynthesis
-
SYSTEMATIC NAME
IUBMB Comments
geranyl-diphosphate diphosphate-lyase (cyclizing, 1,8-cineole-forming)
Requires Mn2+ or Zn2+. Mg2+ is less effective than either. 1,8-Cineole is the main product from the enzyme with just traces of other monoterpenoids. The oxygen atom is derived from water. The reaction proceeds via linalyl diphosphate and alpha-terpineol, the stereochemistry of both depends on the organism. However neither intermediate can substitute for geranyl diphosphate. The reaction in Salvia officinalis (sage) proceeds via (-)-(3R)-linalyl diphosphate [1-3] while that in Arabidopsis (rock cress) proceeds via (+)-(3S)-linalyl diphosphate [4].
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
1,8-cineole cyclase; geranyl pyrophoshate:1,8-cineole cyclase
-
-
-
-
1,8-cineole synthetase
-
-
-
-
plastid terpineol synthase
H2ELN1
-
SabS1
A6XH05
-
SSS
O81193
-
CAS REGISTRY NUMBER
COMMENTARY
110637-19-9
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
-
Q9LDF1
UniProt
Manually annotated by BRENDA team
Eucalyptus sideroxylon
-
-
-
Manually annotated by BRENDA team
parents Lavandula angustifolia and Lavandula latifolia
UniProt
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(3R)-linalyl diphosphate + H2O
? + diphosphate
show the reaction diagram
-
-
-
-
?
(3S)-linalyl diphosphate + H2O
? + diphosphate
show the reaction diagram
-
-
-
-
?
geranyl diphosphate
1,8-cineole + diphosphate
show the reaction diagram
-
-
-
-
?
geranyl diphosphate
1,8-cineole + diphosphate
show the reaction diagram
O81193
-
-
-
?
geranyl diphosphate + H2O
1,8-cineole + diphosphate
show the reaction diagram
Eucalyptus sideroxylon
-
-
-
-
?
geranyl diphosphate + H2O
1,8-cineole + diphosphate
show the reaction diagram
Q9LDF1
-
52% 1,8-cineole plus 0.6% (+)-alpha-thujene, 1.9% (2)-(1S)-alpha-pinene, 14.5% (-)-sabinene, 7.8% (2)-(1S)-beta-pinene, 13.3% myrcene, 4.0% (2)-(4S)-limonene, (E)-2.7% beta-ocimene, 0.8% terpinolene, and 2.4% alpha-terpineol
-
?
geranyl diphosphate + H2O
1,8-cineole + diphosphate
show the reaction diagram
A6XH05, -
-
72.4% 1,8-cineole, 7.1% alpha-terpineol, 9.1% beta-pinene, 4.6% alpha-pinene, 3.6% sabinene, 2.2% myrcene, and <1% limonene
-
?
geranyl diphosphate + H2O
1,8-cineole + diphosphate
show the reaction diagram
O81191
-
79% 1,8-cineole plus 20% of a mixture of (+)- and (-)-alpha-pinene, (+)- and (-)-beta-pinene, myrcene and (+)-sabinene
-
?
geranyl diphosphate + H2O
1,8-cineole + diphosphate
show the reaction diagram
I3WEV8
-
80% 1,8-cineole plus 7.9% sabinene, 6.6% alpha-phellandrene, 2.3% limonene, and 1.7% alpha-terpineol
-
?
geranyl diphosphate + H2O
1,8-cineole + diphosphate
show the reaction diagram
A5Y5L5, -
-
major product is 1,8-cineole plus limonene, sabinene, E-beta-ocimene, b-myrcene, alpha-pinene, and alpha-terpineole
-
?
geranyl diphosphate + H2O
1,8-cineole + diphosphate
show the reaction diagram
H2ELN1
-
products are sabinene, beta-myrcene, limonene, 1,8-cineole, and 36% alpha-terpineol
-
?
geranyl diphosphate + H2O
1,8-cineole + diphosphate
show the reaction diagram
H6WZF2
-
products are sabinene, beta-myrcene, limonene, 1,8-cineole, and 41% alpha-terpineol
-
?
neryl diphosphate + H2O
1,8-cineole + diphosphate
show the reaction diagram
I3WEV8
-
61% 1,8-cineole plus 18.5% sabinene, 8% alpha-phellandrene, 3.3% limonene, and 5.5% alpha-terpineol
-
?
neryl diphosphate + H2O
? + diphosphate
show the reaction diagram
-
-
-
-
?
geranyl diphosphate + H2O
1,8-cineole + diphosphate
show the reaction diagram
-
-
water is the sole source of the ether oxygen atom of 1,8-cineole
-
?
additional information
?
-
Eucalyptus sideroxylon
-
no substrate: farnesyl diphosphate
-
-
-
additional information
?
-
Q9LDF1
no substrate: farnesyl diphosphate
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mg2+
-
or Mn2+, absolutely required. Km value 5 mM
Mg2+
A5Y5L5, -
or Mn2+, required. Mg2+ is preferred over Mn2+
Mg2+
Q9LDF1
or Mn2+, absolutely required. Fully active at 40 mM
Mn2+
-
or Mg2+, absolutely required. Km value 0.3 mM
Mn2+
A5Y5L5, -
or Mg2+, required. Mn2+ is preferred over Mn2+
Mn2+
Q9LDF1
or Mg2+, absolutely required. Fully active at 1 mM
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-Fluorogeranyl diphosphate
-
competitive
2-Fluorolinalyl diphosphate
-
competitive
4-hydroxymercuribenzoate
-
inhibition is reversed by presence of cysteine
benzyl bromide
-
-
diethyl dicarbonate
-
inhibition is reversed by presence of hydroxylamine
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
H6WZF2
enzyme activity reaches its maximum at the second day after anthesis when flowers are fully opened. Enzyme activity is highest at the transition from day to night, diurnal rhythm
-
additional information
H2ELN1
enzyme activity reaches its maximum at the second day after anthesis when flowers are fully opened. Enzyme activity is highest at the transition from day to night, diurnal rhythm
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.002
-
(3R)-linalyl diphosphate
-
pH 6.3, 30C
0.0042
-
(3S)-linalyl diphosphate
-
pH 6.3, 30C
0.0002
-
geranyl diphosphate
Q9LDF1
temperature not specified in the publication, pH not specified in the publication
0.0011
-
geranyl diphosphate
-
pH 6.3, 30C
0.0057
-
geranyl diphosphate
I3WEV8
pH 7.0, 30C
0.007
-
geranyl diphosphate
-
pH 7.1, 31C
0.0119
-
geranyl diphosphate
A6XH05, -
mutant N338A, pH 7.0, 22C
0.0129
-
geranyl diphosphate
A6XH05, -
mutant N338I/A339T/G447S/I449P/P450T, pH 7.0, 22C
0.0144
-
geranyl diphosphate
A6XH05, -
mutant N338S, pH 7.0, 22C
0.0265
-
geranyl diphosphate
A6XH05, -
mutant N338L, pH 7.0, 22C
0.055
-
geranyl diphosphate
A6XH05, -
mutant N338V, pH 7.0, 22C
0.0566
-
geranyl diphosphate
A6XH05, -
mutant N338I, pH 7.0, 22C
0.0654
-
geranyl diphosphate
A6XH05, -
wild-type, pH 7.0, 22C
0.0955
-
geranyl diphosphate
A6XH05, -
mutant N338I/A339T/G447S, pH 7.0, 22C
0.136
-
geranyl diphosphate
A6XH05, -
mutant N338I/A339T, pH 7.0, 22C
0.201
-
geranyl diphosphate
A6XH05, -
mutant N338C, pH 7.0, 22C
0.0045
-
neryl diphosphate
-
pH 6.3, 30C
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.001
-
geranyl diphosphate
Q9LDF1
temperature not specified in the publication, pH not specified in the publication
0.0043
-
geranyl diphosphate
A6XH05, -
mutant N338I/A339T/G447S/I449P/P450T, pH 7.0, 22C; mutant N338I/A339T/G447S, pH 7.0, 22C
0.0088
-
geranyl diphosphate
I3WEV8
pH 7.0, 30C
0.009
-
geranyl diphosphate
A6XH05, -
mutant N338L, pH 7.0, 22C
0.012
-
geranyl diphosphate
A6XH05, -
mutant N338V, pH 7.0, 22C
0.022
-
geranyl diphosphate
A6XH05, -
mutant N338S, pH 7.0, 22C
0.024
-
geranyl diphosphate
A6XH05, -
mutant N338I/A339T, pH 7.0, 22C
0.052
-
geranyl diphosphate
A6XH05, -
mutant N338C, pH 7.0, 22C
0.053
-
geranyl diphosphate
A6XH05, -
wild-type, pH 7.0, 22C
0.069
-
geranyl diphosphate
A6XH05, -
mutant N338A, pH 7.0, 22C
0.096
-
geranyl diphosphate
A6XH05, -
mutant N338I, pH 7.0, 22C
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.045
-
geranyl diphosphate
A6XH05, -
mutant N338I/A339T/G447S, pH 7.0, 22C
10850
0.177
-
geranyl diphosphate
A6XH05, -
mutant N338I/A339T, pH 7.0, 22C
10850
0.22
-
geranyl diphosphate
A6XH05, -
mutant N338V, pH 7.0, 22C
10850
0.26
-
geranyl diphosphate
A6XH05, -
mutant N338C, pH 7.0, 22C
10850
0.33
-
geranyl diphosphate
A6XH05, -
mutant N338I/A339T/G447S/I449P/P450T, pH 7.0, 22C
10850
0.34
-
geranyl diphosphate
A6XH05, -
mutant N338L, pH 7.0, 22C
10850
0.81
-
geranyl diphosphate
A6XH05, -
wild-type, pH 7.0, 22C
10850
1.53
-
geranyl diphosphate
A6XH05, -
mutant N338S, pH 7.0, 22C
10850
1.69
-
geranyl diphosphate
A6XH05, -
mutant N338I, pH 7.0, 22C
10850
5.79
-
geranyl diphosphate
A6XH05, -
mutant N338A, pH 7.0, 22C
10850
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0032
-
2-Fluorogeranyl diphosphate
-
pH 6.3, 30C
0.0048
-
2-Fluorolinalyl diphosphate
-
pH 6.3, 30C
0.005
-
4-hydroxymercuribenzoate
-
pH 6.3, 30C
0.85
-
benzyl bromide
-
pH 6.3, 30C
0.3
-
diethyl dicarbonate
-
pH 6.3, 30C
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8
-
A5Y5L5, -
-
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
35
-
A5Y5L5, -
-
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.6
-
-
isoelectric focusing
5.8
-
-
calculated
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
epidermal gland of leaf
Manually annotated by BRENDA team
Q9LDF1
promoter activity is primarily found in the epidermis, cortex, and stele of mature primary and lateral roots, but not in the root meristem or the elongation zone
Manually annotated by BRENDA team
H6WZF2
pistil
Manually annotated by BRENDA team
O81193
sabinene synthase transcripts show the highest monoterpene synthase transcript expression, about an order of magnitude higher than cineole synthase and borneol synthase
Manually annotated by BRENDA team
-
gene expression of monoterpene synthases and actual monoterpene formation is highest in young leaf
Manually annotated by BRENDA team
H6WZF2
adaxial and abaxial epidermis of petals
Manually annotated by BRENDA team
H2ELN1
adaxial and abaxial epidermis of petals. 1,8-cineole is the major compound emitted
Manually annotated by BRENDA team
A5Y5L5, -
only expressed in sigma/style tissue and in petals
Manually annotated by BRENDA team
Q9LDF1
promoter activity is primarily found in the epidermis, cortex, and stele of mature primary and lateral roots, but not in the root meristem or the elongation zone
Manually annotated by BRENDA team
Q9LDF1
promoter activity is primarily found in the epidermis, cortex, and stele of mature primary and lateral roots, but not in the root meristem or the elongation zone
Manually annotated by BRENDA team
A5Y5L5, -
only expressed in sigma/style tissue and in petals
Manually annotated by BRENDA team
A5Y5L5, -
only expressed in sigma/style tissue and in petals
Manually annotated by BRENDA team
-
glandular trichome
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Eucalyptus sideroxylon
-
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
60000
-
-
gel filtration
72000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
A5Y5L5, -
x * 64550, calculated
?
I3WEV8
x * 63000, SDS-PAGE, x * 68500, calculated
?
Q9LDF1
x * 60000, SDS-PAGE
monomer
-
x * 69400, calculated
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
Eucalyptus sideroxylon
-
sequence contains a putative chloroplast targeting peptide
proteolytic modification
I3WEV8
the N-terminal 52 amino acids are predicted to code for a transit peptide
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
to 1.95 A resolution, space group C2221. alpha-Helical protein comprised of 23 alpha-helices and eight 3/10 helices
A6XH05, -
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0
-
A5Y5L5, -
4 h, loss of about 20% of activity
22
-
A5Y5L5, -
2 h, loss of about 20% of activity
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Glycerol
-
10%, improves stability during the assay
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
Q9LDF1
expression in Escherichia coli
I3WEV8
expression in Escherichia coli
H2ELN1
expression in Escherichia coli
A5Y5L5, -
expression in Escherichia coli
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
LiCINSmRNAlevels parallel the 1,8-cineole content in mature flowers and in developmental stages
I3WEV8
regulation of gene expression by the circadian clock
A5Y5L5, -
daminozide significantly decreases gene expression of the monoterpene synthases sabinene synthase, borneol synthase, and 1,8-cineole synthase
O81193
expression of 1,8-cineole synthase and accumulation of its end product 1,8-cineole remain constant during the growth period until August and then decrease slightly
-
the foliar application of gibberellic acid increases gene expression of the monoterpene synthases sabinene synthase, borneol synthase, and 1,8-cineole synthase. The amounts of two of the endproducts,1,8-cineole and camphor, are directly correlated with the levels of gene expression of the respective monoterpene synthases
O81193
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
N338A
A6XH05, -
products are 11.7% alpha-pinene, 33.1% sabinene, 33.2% beta-pinene, 16.9% myrcene, 5.1% limonene
N338C
A6XH05, -
products are 16.1% alpha-pinene, 41% sabinene, 32.1% beta-pinene, 4.7% myrcene, 6.1% limonene
N338I
A6XH05, -
products are 6.5% alpha-pinene, 48.3% sabinene, 8.2% myrcene, 37% limonene
N338I/A339T
A6XH05, -
products are 5.4% alpha-pinene, 62.1% sabinene, 6.4% myrcene, 26.1% limonene
N338I/A339T/G447S
A6XH05, -
products are 5.7% alpha-pinene, 59.6% sabinene, 5.0% myrcene, 29.7% limonene
N338I/A339T/G447S/I449P/P450T
A6XH05, -
products are 86.8% sabinene, 5.5% myrcene, 7.7% limonene
N338L
A6XH05, -
products are 4.7% alpha-pinene, 5.6% sabinene, 13.2% beta-pinene, 27.2% myrcene, 49.3% limonene
N338S
A6XH05, -
products are 9.8% alpha-pinene, 34.5% sabinene, 34.2% beta-pinene, 6.8% myrcene, 2.6% limonene, 5.6% alpha-cineole, 6.5% alpha-terpineol
additional information
A6XH05, -
conversion of CinS1 to a sabinene synthase based on an amino acid comparison of the enzyme with Salvia officinalis SabS1 and Salvia pomifera SabS1
N338V
A6XH05, -
products are 61.2% sabinene, 8.0% myrcene, 30.8% limonene
additional information
-
exchange of sequences corresponding by homology to the C-terminal domain including the domain spanning alpha-helix, between sabinene synthase and both bornyl diphosphate synthase and cineole synthase. Exchange of residues 304-377 from cineole synthase into sabinene synthase is sufficient to impart the alternative termination chemistry involving water capture to alpha-terpineol and cyclization to 1,8-cineole