Information on EC 4.2.3.16 - (4S)-limonene synthase

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The expected taxonomic range for this enzyme is: Embryophyta

EC NUMBER
COMMENTARY
4.2.3.16
-
RECOMMENDED NAME
GeneOntology No.
(4S)-limonene synthase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
geranyl diphosphate = (S)-limonene + diphosphate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C-O bond cleavage
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
(4R)-carvone biosynthesis
-
-
Biosynthesis of secondary metabolites
-
-
fenchol biosynthesis II
-
-
menthol biosynthesis
-
-
Metabolic pathways
-
-
monoterpene biosynthesis
-
-
Monoterpenoid biosynthesis
-
-
oleoresin monoterpene volatiles biosynthesis
-
-
perillyl aldehyde biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
geranyl-diphosphate diphosphate-lyase [cyclizing; (-)-(4S)-limonene-forming]
A recombinant enzyme (also known as a monoterpene synthase or cyclase) from the grand fir (Abies grandis) requires Mn2+ and K+ for activity. Mg2+ is essentially ineffective as the divalent metal ion cofactor.
CAS REGISTRY NUMBER
COMMENTARY
110639-20-8
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
grand fir
Uniprot
Manually annotated by BRENDA team
grand fir
-
-
Manually annotated by BRENDA team
L. cv. Skunk
-
-
Manually annotated by BRENDA team
Mentha sp.
M. spicata
-
-
Manually annotated by BRENDA team
Mentha sp.
M. x piperita
-
-
Manually annotated by BRENDA team
Mentha sp.
M. x piperita and M. spicata
-
-
Manually annotated by BRENDA team
spearmint
-
-
Manually annotated by BRENDA team
transgenic spike lavender plants expressing the limonene synthase gene from spearmint
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
Bong. Carr.
SwissProt
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(3R)-linalyl diphosphate
?
show the reaction diagram
Mentha sp.
-
-
-
-
?
(3S)-linalyl diphosphate
?
show the reaction diagram
Mentha sp.
-
-
-
-
?
6,7-dihydrogeranyl diphosphate
?
show the reaction diagram
-
-
-
?
6,7-dihydrogeranyl diphosphate
?
show the reaction diagram
-
reaction yields only achiral, olefin products from 6,7-dehydrogeranyl diphosphate, no significant amounts of terpenols or homoterpenols are formed
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
small amounts of (-)-alpha-pinene, (-)-beta-pinene and myrcene as side products
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
small amounts of (-)-alpha-pinene, (-)-beta-pinene and myrcene as side products
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
small amounts of (-)-alpha-pinene, (-)-beta-pinene and myrcene as side products
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
O22340
-
small amounts of (-)-alpha-pinene, (-)-beta-pinene and beta-phellandrene as side products
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
(3S)-linalyl diphosphate as intermediate
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
-
cyclization proceeds via preliminary isomerization to the bound tertiary intermediate 3S-linalyl diphosphate
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
O22340
induced by stem wounding, enhances resistance to insects
small amounts of (-)-alpha-pinene, (-)-beta-pinene and beta-phellandrene as side products
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
essential for menthol production
-
?
geranyl diphosphate
(4S)-limonene + diphosphate
show the reaction diagram
-
-
-
-
?
geranyl diphosphate
limonene + diphosphate
show the reaction diagram
-
step 3 and 4 of monoterpene biosynthesis, geranyl diphosphate is first ionized and isomerized to produce linalyl diphosphate, which either produces the acyclic monoterpenes or the alpha-terpinyl cation, the universal intermediate for cyclic monoterpenes and transformation of the parent structures to various derivatives
limonene is a cyclic monoterpene, minor constituent of the spike lavender oil
-
?
neryl diphosphate
?
show the reaction diagram
Mentha sp.
-
-
-
-
?
geranyl diphosphate
(-)-limonene + diphosphate
show the reaction diagram
Q9M7D0
-
42% terpinolene, 18% (-)-alpha-pinene, 11% (-)-limonene, 10% (-)-beta-pinene plus several minor products
-
?
additional information
?
-
-
overexpression of the LS does not alter the photosynthetic pigment content of spike lavender leaves
-
-
-
additional information
?
-
Q9M7D0
entire product set is derived in stereochemically consistent fashion via (-)-3S-linalyl diphosphate as intermediate
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
-
-
-
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
small amounts of (-)-alpha-pinene, (-)-beta-pinene and myrcene as side products
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
small amounts of (-)-alpha-pinene, (-)-beta-pinene and myrcene as side products
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
-
small amounts of (-)-alpha-pinene, (-)-beta-pinene and myrcene as side products
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
O22340
induced by stem wounding, enhances resistance to insects
small amounts of (-)-alpha-pinene, (-)-beta-pinene and beta-phellandrene as side products
?
geranyl diphosphate
(-)-(4S)-limonene + diphosphate
show the reaction diagram
Mentha sp.
-
essential for menthol production
-
?
geranyl diphosphate
limonene + diphosphate
show the reaction diagram
-
step 3 and 4 of monoterpene biosynthesis, geranyl diphosphate is first ionized and isomerized to produce linalyl diphosphate, which either produces the acyclic monoterpenes or the alpha-terpinyl cation, the universal intermediate for cyclic monoterpenes and transformation of the parent structures to various derivatives
limonene is a cyclic monoterpene, minor constituent of the spike lavender oil
-
?
additional information
?
-
-
overexpression of the LS does not alter the photosynthetic pigment content of spike lavender leaves
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K+
O22340
required, optimum concentration of 0.5 M
Mg2+
O22340
ineffective
Mg2+
Mentha sp.
-
cofactor
Mg2+
Mentha sp.
-
Mn2+ preferred
Mg2+
-
can substitute for Mn2+
Mn2+
O22340
required
Mn2+
Mentha sp.
-
required, optimum activity with 2 mM, inhibitory at higher concentrations
Mn2+
-
required
Mn2+
Mentha sp.
-
required
additional information
Mentha sp.
-
Ca2+, Cd2+, Co2+, Cu2+, Fe2+, Ni2+, Zn2+ activated less than 5%
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
6-cyclopropylidene-(3E)-methyl-hex-2-en-1-yl diphosphate
Mentha sp.
-
Ki: 0.0003 mM, irreversible covalent modification
benzyl bromide
Mentha sp.
-
-
diethyl dicarbonate
Mentha sp.
-
-
diethyl dicarbonate
Mentha sp.
-
reversible with hydroxylamine, pretreatment with geranyl diphosphate and MnCl2 prevents inhibition
linalyl sulfonium ion
Mentha sp.
-
-
N-ethylmaleimide
Mentha sp.
-
reversible by cysteine treatment
p-hydroxymercuribenzoate
Mentha sp.
-
pretreatment with geranyl diphosphate and MnCl2 prevents inhibition
p-hydroxymercuribenzoate
Mentha sp.
-
-
Sodium diphosphate
Mentha sp.
-
Ki: 1 mM
Sodium phosphate
Mentha sp.
-
Ki: 30 mM
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Sodium phosphate
Mentha sp.
-
stimulating at low concentrations
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0019
(3R)-linalyl diphosphate
Mentha sp.
-
-
0.0023
(3S)-linalyl diphosphate
Mentha sp.
-
-
0.00125
geranyl diphosphate
-
-
0.0018
geranyl diphosphate
Mentha sp.
-
-
0.0068
geranyl diphosphate
-
-
0.001
neryl diphosphate
Mentha sp.
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0003
6-cyclopropylidene-(3E)-methyl-hex-2-en-1-yl diphosphate
Mentha sp.
-
irreversible covalent modification
1
Sodium diphosphate
Mentha sp.
-
-
30
Sodium phosphate
Mentha sp.
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
increase in the expression of the spearmint LS cDNA at the different leaf developmental stages is accompanied with increases in the limonene content of transgenic spike lavender lines; leaves of transgenic plants contain increased limonene contents (more than 450% increase compared to controls) that correlates with the highest transcript accumulation of the LS gene.; the effect of the LS expression on oil composition decreases with leaf age as compared to the control; transgenic T0 spike lavender lines overexpressing MsLS show quantitative and qualitative alterations in their terpene profiles, particularly increased amounts of limonene as compared to controls (from 19.05% to 53.97% and from 12.50% to 83.33% in pooled leaves and flowers, respectively). The levels of other monoterpene pathway components are not consistently altered in either leaf or flower oils
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.7
Mentha sp.
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.1 - 7.2
Mentha sp.
-
half maximum activity at both pH
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
Mentha sp.
-
-
Manually annotated by BRENDA team
Mentha sp.
-
glandular trichome secretory cells
Manually annotated by BRENDA team
-
the first two pairs of leaves from 40-day-old seedlings are used as primary explants for transformation
Manually annotated by BRENDA team
-
from natural populations, germinated under sterile conditions
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Mentha sp.
-
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
51000
-
gel filtration
210772
56000
Mentha sp.
-
SDS-PAGE, gel filtration
210765, 210767
56000
Mentha sp.
-
SDS-PAGE
210770, 210773
59800
Mentha sp.
-
calculated from cDNA sequence
210765
70000
-
SDS-PAGE
650491
73500
O22340
calculated MW of recombinant protein expressed in Escherichia coli
210753
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
monomer
Mentha sp.
-
1 * 56000, gel filtration, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
Q9M7D0
sequence contains a putative plastid targeting signal
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure at 2.7 A resolution in two forms liganded to the substrate and intermediate analogs, 2-fluorogeranyl diphosphate and 2-fluorolinalyl diphosphate, respectively
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7
-
most stable
210772
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
unstable in crude secretory cell extract, stable after DEAE-cellulose chromatography
Mentha sp.
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, stable for 1 week without loss of activity after DEAE-cellulose chromatography
Mentha sp.
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
-
Mentha sp.
-
partial
Mentha sp.
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
O22340
expression in Escherichia coli
Q9M7D0
expressed in Escherichia coli
Mentha sp.
-
Agrobacterium-mediated transformation and regeneration of spike lavender is carried out and the transgenic Lavandula latifolia plants express the limonene synthase gene from Mentha spicata. Expression of limonene synthase in transgenic spike lavender results in an altered monoterpene composition in developing leaves, but does not consistently affect essential oil profile in pooled leaves and flowers of transgenic generation T0.
-
expressed in Escherichia coli
-
expression in Escherichia coli
-
expressed in Escherichia coli
-
expressed in Eucalyptus camaldulensis
-
expression in Escherichia coli
Q675L1
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the expression of (4S)-limonene synthase, 2-C-methyl-D-erythritol-2,4-cyclodiphosphate synthase Mds, (+)-pulegone reductase Pr, and (+)-menthofuran synthase Mfs do not show significant changes upon UV-B irradioation of field-grown and growth chamber plants
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
exchange of the predicted helix D through F region gives rise to an (-)-limonene/(-)-alpha-pinene synthase product outcome
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
agriculture
-
putative production of transgenic aromatic plants overexpressing monoterpene synthases
industry
-
limonene is an aromatic component in numerous products like beverages and cosmetics
synthesis
-
engineering of Echerichia coli with a heterologous mevalonate pathway and limonene synthase for production of limonene followed by coupling with a cytochrome P450,which specifically hydroxylates limonene to produce perillyl alcohol. A strain containing all mevalonate pathway genes in a single plasmid produces limonene at titers over 400 mg/l from glucose. Incorporation of a cytochrome P450 to hydroxylate limonene yields approximately 100mg/l of perilyl alcohol
agriculture
-
UV-B irradiation differentially modulates the expression of genes involved in peppermint essential oil biogenesis and the content of UV-B absorbing flavonoids. Plants grown in field are better adapted to increasing UV-B irradiation than plants cultivated in growth chambers