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
-
(+)-pinene cyclase
-
-
(-)-pinene cyclase
-
-
ag9
-
-
-
-
LaLIMS_RR
-
-
-
-
limonene synthase
-
PmeTPS2
-
-
-
-
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
-
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
-
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
-
wild-type produces 1.3% (+)-car-3-ene, 44.7% (-)-sabinene, 35.9% terpinolene
?
geranyl diphosphate
terpinolene + diphosphate
show the reaction diagram
-
42% terpinolene, 18% (-)-alpha-pinene, 11% (-)-limonene, 10% (-)-beta-pinene plus several minor products
?
geranyl diphosphate
terpinolene + diphosphate
show the reaction diagram
-
78% (+)-car-3-ene plus 11% terpinolene plus minor products
?
geranyl diphosphate
terpinolene + diphosphate
show the reaction diagram
-
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
-
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
-
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
?
-
entire product set is derived in stereochemically consistent fashion via (-)-3S-linalyl diphosphate as intermediate
-
-
-
additional information
?
-
no oxygenated or phosphorylated monoterpenes are found in detectable amounts
-
-
-
additional information
?
-
no substrates: farnesyl diphosphate, geranygeranyl diphosphate
-
-
-
additional information
?
-
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+
or Mn2+, reuired. Optimum concentraion 50 mM
Mn2+
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
mutant L596F, 30C, pH not specified in the publication
0.0158
geranyl diphosphate
wild-type, 30C, pH not specified in the publication
0.0202
geranyl diphosphate
mutant A595G/F596L/L599F, 30C, pH not specified in the publication
0.0316
geranyl diphosphate
mutant F596L, 30C, pH not specified in the publication
0.0357
geranyl diphosphate
wild-type, 30C, pH not specified in the publication
0.0474
geranyl diphosphate
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
Picea sitchensis
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant A595G/F596L/L599F, 30C, pH not specified in the publication
0.0825
geranyl diphosphate
Picea sitchensis
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant F596L, 30C, pH not specified in the publication
0.098
geranyl diphosphate
Picea sitchensis
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
wild-type, 30C, pH not specified in the publication
0.545
geranyl diphosphate
Picea sitchensis
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
wild-type, 30C, pH not specified in the publication
0.643
geranyl diphosphate
Picea sitchensis
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
Picea sitchensis
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant A595G/F596L/L599F, 30C, pH not specified in the publication
175
0.0027
geranyl diphosphate
Picea sitchensis
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant F596L, 30C, pH not specified in the publication
175
0.0062
geranyl diphosphate
Picea sitchensis
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
wild-type, 30C, pH not specified in the publication
175
0.0153
geranyl diphosphate
Picea sitchensis
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
wild-type, 30C, pH not specified in the publication
175
0.042
geranyl diphosphate
Picea sitchensis
F1CKI6, F1CKI8, F1CKI9, F1CKJ1
mutant L596F, 30C, pH not specified in the publication
175
0.255
geranyl diphosphate
Lavandula angustifolia
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
calculated
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
x * 70300, SDS-PAGE
?
x * 71912, calculated
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
sequence contains a putative plastid targeting signal
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
expression in Escherichia coli
expression in Escherichia coli
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
treatment with methyl jasmonate induces transient transcript accumulation in stem tissues
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A595G
shift in product profile, mutant produces 2.4% (+)-car-3-ene, 39% (-)-sabinene, 33.3% terpinolene
A595G/F596L/L599F
shift in product profile, mutant produces 42.3% (+)-car-3-ene, 7.3% (-)-sabinene, 20.1% terpinolene
F596E
shift in product profile, mutant produces 0.5% (+)-car-3-ene, 10.2% (-)-sabinene, 5.7% terpinolene, 70.9% limonene
F596G
shift in product profile, mutant produces 10.0% (+)-car-3-ene, 11.2% (-)-sabinene, 17.3% terpinolene, 27.0% myrcene, 17.0% limonene
F596H
shift in product profile, mutant produces 2.1% (+)-car-3-ene, 46.5% (-)-sabinene, 27.1% terpinolene
F596L
shift in product profile, mutant produces 28.6% (+)-car-3-ene, 42.8% (-)-sabinene, 36.4% terpinolene
G595A/L596F/F599L
shift in product profile, mutant produces 1.8% (+)-car-3-ene, 23.3% (-)-sabinene, 55.2% terpinolene; shift in product profile, mutant produces 4.7% (+)-car-3-ene, 47.4% (-)-sabinene, 35.2% terpinolene; shift in product profile, mutant produces 5.4% (+)-car-3-ene, 29.0% (-)-sabinene, 26.4% terpinolene
L596F
shift in product profile, mutant produces 12.3% (+)-car-3-ene, 37.4% (-)-sabinene, 35.4% terpinolene; shift in product profile, mutant produces 5% (+)-car-3-ene, 20.9% (-)-sabinene, 53.5% terpinolene; shift in product profile, mutant produces 9.2% (+)-car-3-ene, 20.2% (-)-sabinene, 32.2% terpinolene
S589A
shift in product profile, mutant produces 1.2% (+)-car-3-ene, 42.9% (-)-sabinene, 35.5% terpinolene
L599F
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; 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; 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; 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