Information on EC 4.2.3.113 - terpinolene synthase

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The expected taxonomic range for this enzyme is: Spermatophyta

EC NUMBER
COMMENTARY
4.2.3.113
-
RECOMMENDED NAME
GeneOntology No.
terpinolene synthase
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
geranyl diphosphate = terpinolene + diphosphate
show the reaction diagram
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
monoterpene biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
geranyl-diphosphate diphosphate-lyase (cyclizing, terpinolene-forming)
Requires Mg2+. Mn2+ is less effective and product ratio changes. Forms traces of other monoterpenoids.
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
(+)-3-carene synthase
F1CKI6, F1CKI8, F1CKI9
-
(+)-pinene cyclase
-
-
(-)-pinene cyclase
-
-
ag9
-
-
-
-
ag9
Q9M7D0
-
JF67
Q84SM8
-
LaLIMS_RR
-
-
-
-
limonene synthase
Q2XSC6
-
PmeTPS2
-
-
-
-
TPS-3car1
F1CKI6
-
TPS-3car2
F1CKI8
-
TPS-3car3
F1CKI9
-
TPS-sab
F1CKJ1
-
CAS REGISTRY NUMBER
COMMENTARY
250376-08-0
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
isoform TPS-3car1
UniProt
Manually annotated by BRENDA team
isoform TPS-3car2
UniProt
Manually annotated by BRENDA team
isoform TPS-3car3
UniProt
Manually annotated by BRENDA team
isoform TPS-sab
UniProt
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
terpinolene + diphosphate
show the reaction diagram
-
-
products are 17% (-)-alpha-pinene, 9% beta-pinene, 3% camphene, 28% limonene, 9% terpinolene, 23% myrcene, 3% cis-ocimene, 13% trans-ocimene, products are 62% (+)-alpha-pinene, 6% camphene, 11% limonene, 7% terpinolene, 10% myrcene, 1% cis-ocimene, 4% trans-ocimene
-
?
(3S)-linalyl diphosphate
terpinolene + diphosphate
show the reaction diagram
-
-
products are 15% (-)-alpha-pinene, 18% beta-pinene, 12% camphene, 11% limonene, 6% terpinolene, 23% myrcene, 3% cis-ocimene, 8% trans-ocimene, products are 9% (+)-alpha-pinene, 9% camphene, 34% limonene, 28%% terpinolene, 11% myrcene, 2% cis-ocimene, 6% trans-ocimene
-
?
2 geranyl diphosphate
(+)-car-3-ene + terpinolene + 2 diphosphate
show the reaction diagram
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
-
wild-type produces 46.2% (+)-car-3-ene, 8.8% (-)-sabinene, 29.7% terpinolene
-
?
2 geranyl diphosphate
(+)-car-3-ene + terpinolene + 2 diphosphate
show the reaction diagram
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
-
wild-type produces 67.5% (+)-car-3-ene, 6.9% (-)-sabinene, 15.4% terpinolene
-
?
2 geranyl diphosphate
(-)-sabinene + terpinolene + 2 diphosphate
show the reaction diagram
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
-
wild-type produces 1.3% (+)-car-3-ene, 44.7% (-)-sabinene, 35.9% terpinolene
-
?
geranyl diphosphate
terpinolene + diphosphate
show the reaction diagram
Q9M7D0
-
42% terpinolene, 18% (-)-alpha-pinene, 11% (-)-limonene, 10% (-)-beta-pinene plus several minor products
-
?
geranyl diphosphate
terpinolene + diphosphate
show the reaction diagram
Q84SM8
-
78% (+)-car-3-ene plus 11% terpinolene plus minor products
-
?
geranyl diphosphate
terpinolene + diphosphate
show the reaction diagram
Q4QSN6
-
main product terpinolene, plus minor products (+)-alpha-pinene, gamma-terpinene, alpha-terpinene, (-)-limonene, sabinene, (-)-beta-pinene, and 3-carene
-
?
geranyl diphosphate
terpinolene + diphosphate
show the reaction diagram
-
-
products are 28% (-)-alpha-pinene, 35% beta-pinene, 24% camphene, 5% limonene, 2% terpinolene, 6% myrcene, products are 49% (+)-alpha-pinene, 30% camphene, 10% limonene, 5% terpinolene, 6% myrcene
-
?
geranyl diphosphate
terpinolene + diphosphate
show the reaction diagram
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
-
wild-type produces 49% (+)-car-3-ene, 8.7% (-)-sabinene, 24.7% terpinolene
-
?
neryl diphosphate
terpinolene + diphosphate
show the reaction diagram
-
-
products are 14% (+)-alpha-pinene, 9% camphene, 62% limonene, 8% terpinolene, 6% myrcene, products are 18% (-)-alpha-pinene, 18% beta-pinene, 12% camphene, 42% limonene, 9% terpinolene, 9% myrcene
-
?
geranyl diphosphate
terpinolene
show the reaction diagram
Q2XSC6
-
products are 39% (R)-(+)-limonene, 22% terpinolene, 16% (1R,5S)-(+)-camphene, 14% (1R,5R)-(+)-alpha-pinene, 8% betamyrcene and traces of alpha-phellandrene. Product pattern changes markedly to 46% limonene, 9% terpinolene, 23% alpha-pinene, 5% beta-myrcene and 4% a-phellandrene, when Mn2+ is supplied instead of Mg2+
-
?
additional information
?
-
Q9M7D0
entire product set is derived in stereochemically consistent fashion via (-)-3S-linalyl diphosphate as intermediate
-
-
-
additional information
?
-
Q84SM8
no oxygenated or phosphorylated monoterpenes are found in detectable amounts
-
-
-
additional information
?
-
Q4QSN6
no substrates: farnesyl diphosphate, geranygeranyl diphosphate
-
-
-
additional information
?
-
Q2XSC6
no substrates: farnesyl diphosphate, geranylgeranyl diphosphate
-
-
-
additional information
?
-
-
reaction follows a cisoid, anti-endo-pattern. In the case of geranyl diphosphate, a preassociation mechanism is suggested in which optimum folding of the terpenyl chain precedes the initial ionization step. The alternate substratse are ionized by the cyclases prior to their achieving the optimum orientation for bicyclization
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Q2XSC6
or Mn2+, reuired. Optimum concentraion 50 mM
Mn2+
Q2XSC6
or Mg2+, required. Optimum concentration 0.2 mM, 68% of the efficieny with Mg2+. Product pattern changes markedly to 46% limonene, 9% terpinolene, 23% alpha-pinene, 5% beta-myrcene and 4% alpha-phellandrene, when Mn2+ is supplied instead of Mg2+
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0035
(3R)-linalyl diphosphate
-
pH 6.5, 30C
0.004
(3R)-linalyl diphosphate
-
pH 6.5, 30C
0.0035
(3S)-linalyl diphosphate
-
pH 6.5, 30C
0.0049
(3S)-linalyl diphosphate
-
pH 6.5, 30C
0.0021
geranyl diphosphate
-
pH 6.5, 30C
0.003
geranyl diphosphate
-
pH 6.5, 30C
0.0153
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant L596F, 30C, pH not specified in the publication
0.0158
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
wild-type, 30C, pH not specified in the publication
0.0202
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant A595G/F596L/L599F, 30C, pH not specified in the publication
0.0316
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant F596L, 30C, pH not specified in the publication
0.0357
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
wild-type, 30C, pH not specified in the publication
0.0474
geranyl diphosphate
Q2XSC6
pH 7.0, 30C
0.0021
neryl diphosphate
-
pH 6.5, 30C
0.006
neryl diphosphate
-
pH 6.5, 30C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0352
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant A595G/F596L/L599F, 30C, pH not specified in the publication
0.0825
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant F596L, 30C, pH not specified in the publication
0.098
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
wild-type, 30C, pH not specified in the publication
0.545
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
wild-type, 30C, pH not specified in the publication
0.643
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant L596F, 30C, pH not specified in the publication
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0017
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant A595G/F596L/L599F, 30C, pH not specified in the publication
175
0.0027
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant F596L, 30C, pH not specified in the publication
175
0.0062
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
wild-type, 30C, pH not specified in the publication
175
0.0153
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
wild-type, 30C, pH not specified in the publication
175
0.042
geranyl diphosphate
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant L596F, 30C, pH not specified in the publication
175
0.255
geranyl diphosphate
Q2XSC6
pH 7.0, 30C
175
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.2
Q84SM8
calculated
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
Q2XSC6
x * 70300, SDS-PAGE
?
Q84SM8
x * 71912, calculated
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
Q9M7D0
sequence contains a putative plastid targeting signal
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
Q9M7D0
expression in Escherichia coli
Q2XSC6
expression in Escherichia coli
Q84SM8
-
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
treatment with methyl jasmonate induces transient transcript accumulation in stem tissues
Q84SM8
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A595G
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 2.4% (+)-car-3-ene, 39% (-)-sabinene, 33.3% terpinolene
A595G/F596L/L599F
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 42.3% (+)-car-3-ene, 7.3% (-)-sabinene, 20.1% terpinolene
F596E
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 0.5% (+)-car-3-ene, 10.2% (-)-sabinene, 5.7% terpinolene, 70.9% limonene
F596G
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 10.0% (+)-car-3-ene, 11.2% (-)-sabinene, 17.3% terpinolene, 27.0% myrcene, 17.0% limonene
F596H
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 2.1% (+)-car-3-ene, 46.5% (-)-sabinene, 27.1% terpinolene
F596L
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 28.6% (+)-car-3-ene, 42.8% (-)-sabinene, 36.4% terpinolene
F596R
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
inactive
G595A/L596F/F599L
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 1.8% (+)-car-3-ene, 23.3% (-)-sabinene, 55.2% terpinolene
G595A/L596F/F599L
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 4.7% (+)-car-3-ene, 47.4% (-)-sabinene, 35.2% terpinolene
G595A/L596F/F599L
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 5.4% (+)-car-3-ene, 29.0% (-)-sabinene, 26.4% terpinolene
L596F
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 12.3% (+)-car-3-ene, 37.4% (-)-sabinene, 35.4% terpinolene
L596F
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 5% (+)-car-3-ene, 20.9% (-)-sabinene, 53.5% terpinolene
L596F
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 9.2% (+)-car-3-ene, 20.2% (-)-sabinene, 32.2% terpinolene
S589A
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 1.2% (+)-car-3-ene, 42.9% (-)-sabinene, 35.5% terpinolene
L599F
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
shift in product profile, mutant produces 1.5% (+)-car-3-ene, 20.2% (-)-sabinene, 32.2% terpinolene
additional information
-
construction of a series of domain swaps and site-directed substitutions between (-)-sabinene synthase and (+)-3-carene synthase isoforms to explore which regions and specific amino acids of these enzymes affect the differences of their product profiles. The amino acid in position 596 is critical for product profiles dominated by (+)-3-carene in (+)-3-carene synthase isoforms. A leucine in this position promotes formation of (+)-3-carene, whereas phenylalanine promotes (-)-sabinene
additional information
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
construction of a series of domain swaps and site-directed substitutions between (-)-sabinene synthase and (+)-3-carene synthase isoforms to explore which regions and specific amino acids of these enzymes affect the differences of their product profiles. The amino acid in position 596 is critical for product profiles dominated by or (-)-sabinene. A leucine in this position promotes formation of (+)-3-carene, whereas phenylalanine promotes (-)-sabinene