Information on EC 4.2.3.120 - (-)-beta-pinene synthase

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The expected taxonomic range for this enzyme is: Spermatophyta

EC NUMBER
COMMENTARY
4.2.3.120
-
RECOMMENDED NAME
GeneOntology No.
(-)-beta-pinene synthase
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
geranyl diphosphate = (-)-beta-pinene + diphosphate
show the reaction diagram
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
monoterpene biosynthesis
-
Monoterpenoid biosynthesis
-
oleoresin monoterpene volatiles biosynthesis
-
SYSTEMATIC NAME
IUBMB Comments
geranyl-diphosphate diphosphate-lyase [cyclizing, (-)-beta-pinene-forming]
Cyclase II of Salvia officinalis (sage) produces about equal parts (-)-alpha-pinene, (-)-beta-pinene and (-)-camphene, plus traces of other monoterpenoids. The enzyme, which requires Mg2+ (preferred to Mn2+), can also use (3S)-Linalyl diphosphate (preferred to (3R)-linalyl diphosphate) [1-4]. The enzyme from Abies grandis (grand fir) produces roughly equal parts of (-)-alpha-pinene and (-)-beta-pinene [6-9]. Cyclase IV from Pinus contorta (lodgepole pine) produces 63% (-)-beta-pinene, 26% 3-carene, and traces of alpha-pinene [10]. Synthase III from Pinus taeda (loblolly pine) forms (-)-beta-pinene with traces of alpha-pinene and requires Mn2+ and K+ (Mg2+ is ineffective) [11]. A cloned enzyme from Artemisia annua (sweet wormwood) gave (-)-beta-pinene with traces of (-)-alpha-pinene [5]. The enzyme from Picea sitchensis (Sika spruce) forms 30% (-)-beta-pinene and 70% (-)-alpha-pinene [12]. See also EC 4.2.3.119, (-)-alpha-pinene synthase, EC 4.2.3.117, (-)-camphene synthase, and EC 4.2.3.107 (+)-3-carene synthase.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
(-)-(1S,5S)-pinene synthase
-
-
-
-
(-)-pinene synthase
O24475
-
ag9
Q9M7D0
-
beta-geraniolene synthase
-
-
-
-
mono-tps
Q6XDB5
-
QH6
Q94G53
-
TPS2
Q6XDB5
-
CAS REGISTRY NUMBER
COMMENTARY
110637-20-2
for both EC 4.2.3.119 and 4.2.3.120
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
isoform VI
-
-
Manually annotated by BRENDA team
isoform cyclase IV
-
-
Manually annotated by BRENDA team
isoform synthase III
-
-
Manually annotated by BRENDA team
isoform cyclase II
-
-
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
(-)-beta-pinene + diphosphate
show the reaction diagram
-
-
products are (-)-alpha-pinene, (-)-beta-pinene, and (-)-camphene
-
?
(3R)-linalyl diphosphate
myrcene + diphosphate
show the reaction diagram
-
-
-
-
?
(3S)-linalyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
-
-
63% (-)-beta-pinene, 25.8% (+-)carene, and 6.7% alpha-pinene in almost racemic mixture.Products are exactly the same as with geranyl diphosphate
-
?
(3S)-linalyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
-
-
products are (-)-alpha-pinene, (-)-beta-pinene, and (-)-camphene
-
?
geranyl diphoshate
(-)-beta-pinene + diphosphate
show the reaction diagram
Q9M7D0
-
42% terpinolene, 18% (-)-alpha-pinene, 11% (-)-limonene, 10% (-)-beta-pinene plus several minor products
-
?
geranyl diphosphate
(+)-beta-pinene + diphosphate
show the reaction diagram
-
-
main product
-
?
geranyl diphosphate
(-)-beta-pinene
show the reaction diagram
-
-
products are 28% (-)-alpha-pinene, 35% (-)-beta-pinene, 24% (+)-camphene, 5% (+)-limonene, plus some terpinolene and myrcene
-
?
geranyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
-
-
25% (-)-alpha-pinene, 31% (-)-camphene, 24% (-)beta-pinene
-
?
geranyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
-
-
63% (-)-beta-pinene, 25.8% (+-)carene, and 6.7% alpha-pinene in almost racemic mixture. (+)-alpha-pinene arises from the rare isomerization of geranyl diphosphate to (3R)-linalyl diphosphate
-
?
geranyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
-
-
about 40% (-)-alpha-pinene and 60% (-)-beta-pinene
-
?
geranyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
-
-
main products are (-)-alpha-pinene, (-)-beta-pinene and camphene. Primary deuterium isotope effects suggest that (-)-alpha-pinene and (-)-beta-pinene derive from alternative deprotonation of a common enzymatic intermediate
-
?
geranyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
-, O24475
-
products are (-)-alpha-pinene and (-)-beta pinene
-
?
geranyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
Q94G53
-
products are (-)-alpha-pinene and (-)-beta-pinene, in a ratio of 6:94
-
?
geranyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
Q6XDB5
-
products are (-)-alpha-pinene and (-)-beta-pinene, in a ratio of about 35:10
-
?
geranyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
-
-
products are (-)-alpha-pinene, (-)-beta-pinene, and (-)-camphene
-
?
geranyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
-
-
products are (-)-camphene, (-)-alpha-pinene, (-)-beta-pinene, (-)-limonene and myrcene
-
?
geranyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
-
-
products are 29% (-)-alpha-pinene, 63% (-)-beta-pinene, 1.8% myrcene, 3.6% limonene
-
?
geranyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
-
-
products are 31.9% (-)-alpha-pinene and 63.9% (-)-beta-pinene, plus 4.2% myrcene
-
?
neryl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
-
-
products are (-)-alpha-pinene, (-)-beta-pinene, and (-)-camphene
-
?
geranyl diphosphate
(-)-beta-pinene + diphosphate
show the reaction diagram
Q8L5K2
-
products are beta-pinene (81.4%, almost exclusively (-)-beta-pinene), sabinene (11%), alpha-pinene (4.1%), limonene (3.5%) and a trace of gamma-terpinene
-
?
additional information
?
-
-
bicyclic products pinene and camphene are derived via the cyclization of the bound, tertiary intermediate (3R)-linalyl diphosphate. Limonene is formed via conformational foldings in addition to the 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
-
-
-
additional information
?
-
-
each product exhibits the same absolute configuration at the center derived from C-6 of geranyl diphosphate, i e. the isopropylidene-substituted carbon
-
-
-
additional information
?
-
Q9M7D0
entire product set is derived in stereochemically consistent fashion via (-)-3S-linalyl diphosphate as intermediate
-
-
-
additional information
?
-
-
enzymes removes the C4-proS-hydrogen of the substrate, the C3 proton of the corresponding pinyl cation, with a stereoselectivity exceeding 78% in the formation of (-)-alpha-pinene
-
-
-
additional information
?
-
-
product distribution varies with deuterium substitution at C4 and C10 of substrate. Kinetic isotope effects strongly indicate multiple bicyclic olefin production through the partitioning of common carbocation intermediates
-
-
-
additional information
?
-
-
substrates geranyl, neryl, and (3S)-linalyl diphosphate yield exclusively the (-)-isomer series, whereas (3R)-linalyl diphosphate affords the (+)-isomers at low rates
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
K+
-
required, Km value 7.5 mM
Mg2+
-
very poor substitute for Mn2+
Mg2+
Q8L5K2
or Mn2+, Mn2+ is preferred over Mg2+
Mg2+
-
or Mn2+, required. Mg2+ is preferred over Mn2+
Mn2+
-
absolutely required, Km value 0.03 mM
Mn2+
Q8L5K2
or Mg2+, Mn2+ is preferred over Mg2+, optimum concentration 0.6 mM
Mn2+
-
absolutely required
Mn2+
-
or Mn2+, required. Mg2+ is preferred over Mn2+, Mn2+ shows 60-70% of the activity with Mg2+
additional information
-
Cu2+, Cd2+, Co2+, Ni2+ and Zn2+ are ineffective
additional information
-
not effective: Li+, Na+, Rb+, Cs+
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
4-hydroxymercuribenzoate
-
-
4-hydroxymercuribenzoate
-
-
alpha-pinene
-
-
diethyldicarbonate
-
-
diphosphate
-
-
N-ethylmaleimide
-
-
Phenylglyoxal
-
inactivation, coincubation with substrate and metal-ion cofactor reduces the rate of inactivation by 10-fold
phosphate
-
-
geranyl diphosphate
-
substrate inhibition above 0.05 mM
additional information
-
enzyme is inactivated by thiol-modifying agents, coincubation with substrate and metal-ion cofactor does not protect
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
activity increases upon stem wounding
-
additional information
-
enzyme activity increases upon wounding
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0032
-
(3R)-linalyl diphosphate
-
pH 7.0, 31C
0.0036
-
(3R)-linalyl diphosphate
-
pH 7.2, 30C
0.001
-
(3S)-linalyl diphosphate
-
pH 7.0, 31C
0.0036
-
(3S)-linalyl diphosphate
-
pH 7.2, 30C
0.0025
-
geranyl diphosphate
-
pH 7.0, 31C
0.003
-
geranyl diphosphate
-
pH 6.5, 30C
0.0031
-
geranyl diphosphate
-
pH 7.2, 30C
0.0031
-
geranyl diphosphate
Q8L5K2
pH 7.0, 30C
0.006
-
geranyl diphosphate
-
pH 7.8, temperature not specified in the publication
0.0023
-
neryl diphosphate
-
pH 7.2, 30C
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0019
-
4-hydroxymercuribenzoate
-
pH 7.8, temperature not specified in the publication
0.64
-
diethyldicarbonate
-
pH 7.8, temperature not specified in the publication
0.17
-
diphosphate
-
pH 7.8, temperature not specified in the publication
51
-
phosphate
-
pH 7.8, temperature not specified in the publication
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.1
-
-
-
7.8
-
-
-
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.7
-
-
half-maximal activity
7
-
-
80% of maximum activity
7.9
-
-
half-maximal activity
8.3
-
-
80% of maximum activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.5
-
-
or below, isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Q8L5K2
glants of fruit lavedo and fruit peel
Manually annotated by BRENDA team
Q94G53
juvenile leaf
Manually annotated by BRENDA team
-
immature leaf
Manually annotated by BRENDA team
-
activity increases upon stem wounding
Manually annotated by BRENDA team
-
sapling stem
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
63000
-
-
gel filtration
65000
-
-
gel fitration
95000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 71500, calculated
?
-
x * 62000, SDS-PAGE
monomer
-
1 * 62000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
-
sequence contains a putative N-terminal plastid targeting signal
proteolytic modification
Q9M7D0
sequence contains a putative plastid targeting signal
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
modeling of amino acid sequence onto the crystal structures of tobacco 5-epi-aristolochene synthase and bornyl diphosphate synthase and comparison with (-)-camphene synthase
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
purified enzyme, stable at 4C, in 50 mM Hepes, pH 7.8, for at least 3 weeks, at -20C, stable for several months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
partial
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
-
expression in Escherichia coli
Q8L5K2
expression in Escherichia coli
Q6XDB5
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
transcript levels are transiently reduced following mechanical wounding or fungal elicitor treatment
Q94G53
under a photoperiod of 12 h/12 h (light/dark), the abundance of QH6 transcripts fluctuates in a diurnal pattern that ebbs around 3 h before daybreak, i.e. 9th h in the dark phase, and peaks after 9 h in light, i.e. 9th h in the light phase. The contents of (-)--pinene in juvenile leaves and in emitted volatiles also vary in a diurnal rhythm, correlating strongly with mRNA accumulation
Q94G53
significant increase in transcript level in the leaders of lateral branches of weevil-attacked and mechanically wounded trees
Q6XDB5
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C372S
-
replacement with corresponding residue of (-)-camphene synthase, 97% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene, while the levels of total pinenes remains relatively constant
C372S/C480S
-
replacement with corresponding residue of (-)-camphene synthase, 72% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
C372S/F597W
-
replacement with corresponding residue of (-)-camphene synthase, 100% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
C372S/F597W/S485C/F597W
-
replacement with corresponding residue of (-)-camphene synthase, 99% of wild-type activity. Mutant produces about 80%(-)-alpha-pinene and 10% (-)-beta-pinene
C372S/S485C
-
replacement with corresponding residue of (-)-camphene synthase, 92% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
C480S
-
replacement with corresponding residue of (-)-camphene synthase, 97% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene, while the levels of total pinenes remains relatively constant
C480S/F597W
-
replacement with corresponding residue of (-)-camphene synthase, 7% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
C480S/S485C
-
replacement with corresponding residue of (-)-camphene synthase, 70% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
S485C
-
replacement with corresponding residue of (-)-camphene synthase, 100% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene, while the levels of total pinenes remains relatively constant
S485C/F597W
-
replacement with corresponding residue of (-)-camphene synthase, 68% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene
F597W
-
replacement with corresponding residue of (-)-camphene synthase, 73% of wild-type activity. Mutant produces an increased proportion of (-)-alpha-pinene and a correspondingly decreased proportion of (-)-beta-pinene, while the levels of total pinenes remains relatively constant
additional information
-
replacement of selected amino acid residues in (-)-pinene synthase with the corresponding residues from (-)-camphene synthase in an effort to identify the amino acids responsible for the catalytic diVerences. The approach produces an enzyme in which more than half of the product is channeled through an alternative pathway. Several (-)-pinene synthase to (-)-camphene synthase amino acid substitutions are necessary before catalysis is significantly altered
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
renaturation after SDS-PAGE in 1% Tween 20
-