Information on EC 1.21.3.7 - tetrahydrocannabinolic acid synthase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Cannabis sativa

EC NUMBER
COMMENTARY
1.21.3.7
-
RECOMMENDED NAME
GeneOntology No.
tetrahydrocannabinolic acid synthase
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
cannabigerolate + O2 = DELTA9-tetrahydrocannabinolate + H2O2
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
oxidative cyclization
PATHWAY
KEGG Link
MetaCyc Link
cannabinoid biosynthesis
-
SYSTEMATIC NAME
IUBMB Comments
cannabigerolate:oxygen oxidoreductase (cyclizing, DELTA9-tetrahydrocannabinolate-forming)
A flavoprotein (FAD). The cofactor is covalently bound. Part of the cannabinoids biosynthetic pathway in the plant Cannabis sativa. The enzyme can also convert cannabinerolate (the (Z)-isomer of cannabigerolate) to Delta9-THCA with lower efficiency. Whereas the product was originally called Delta1-tetrahydrocannabinolate, the recommended name according to systematic peripheral numbering is Delta9-tetrahydrocannabinolate.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
DELA1-THCA synthase
-
-
DELTA1-tetrahydrocannabinolic acid synthase
-
-
-
-
DELTA1-tetrahydrocannabinolic acid synthase
-
-
DELTA1-tetrahydrocannabinolic acid synthase
Q8GTB6
-
tetrahydrocannabinolic acid synthase
-
-
tetrahydrocannabinolic-acid synthase
-
-
THCA synthase
-
-
-
-
THCA synthase
Q8GTB6
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Mexican strain
-
-
Manually annotated by BRENDA team
Mexican strain
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
metabolism
-
key enzyme in the synthesis of tetrahydrocannabinolate
physiological function
-
a key enzyme controlling the psychoactivity of cannabis
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
cannabigerolate + O2
DELTA9-tetrahydrocannabinolate + H2O2
show the reaction diagram
-
-
-
-
?
cannabigerolate + O2
DELTA9-tetrahydrocannabinolate + H2O2
show the reaction diagram
-
-
-
-
ir
cannabigerolate + O2
DELTA9-tetrahydrocannabinolate + H2O2
show the reaction diagram
Q8GTB6
-
-
-
?
cannabigerolate + O2
DELTA9-tetrahydrocannabinolate + H2O2
show the reaction diagram
-, Q8GTB6
-
-
-
ir
cannabigerolate + O2
DELTA9-tetrahydrocannabinolate + H2O2
show the reaction diagram
-
best substrate
-
-
?
cannabigerolate + O2
DELTA9-tetrahydrocannabinolate + H2O2
show the reaction diagram
-
about 98% conversion rate in 24 h
-
-
?
cannabigerolate + O2
DELTA9-tetrahydrocannabinolate + H2O2
show the reaction diagram
-
maximum conversion rate of about 98% in 24 h
-
-
?
cannabinerolate + O2
DELTA9-tetrahydrocannabinolate + H2O2
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
cannabigerol and cannabinero are no substrates
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
FAD
-
the enzyme contains covalently attached FAD at a molar ratio of FAD to protein of 1:1
FAD
-, Q8GTB6
the enzyme contains covalently attached FAD cofactor at a molar ratio of FAD to protein of 1:1
additional information
-
the enzyme does not require any cofactors or coenzymes
-
additional information
-
NAD+, NADP+, FAD, and FMN do not stimulate the enzyme activity
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
the enzyme does not require any metal ions for activity
additional information
-
the enzyme is not influenced by Ca2+, Mg2+ and Mn2+
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Fe3+
-
91% residual activity at 2 mM Fe3+
-
Hg2+
-
strong inhibitor, 4% residual activity at 2 mM Hg2+
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.131
-
cannabigerolate
-
recombinant enzyme, in 100 mM sodium citrate (pH 5.5), at 30C
0.134
-
cannabigerolate
-
at pH 6.0, temperature not specified in the publication
0.446
-
cannabigerolate
-, Q8GTB6
mutant enzyme R108A, in 100 mM sodium citrate buffer (pH 5.0), at 30C
0.54
-
cannabigerolate
-, Q8GTB6
wild type enzyme, in 100 mM sodium citrate buffer (pH 5.0), at 30C
0.783
-
cannabigerolate
-, Q8GTB6
mutant enzyme R110A, in 100 mM sodium citrate buffer (pH 5.0), at 30C
0.254
-
cannabinerolate
-
at pH 6.0, temperature not specified in the publication
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.043
-
cannabigerolate
-, Q8GTB6
mutant enzyme R108A, in 100 mM sodium citrate buffer (pH 5.0), at 30C
0.075
-
cannabigerolate
-, Q8GTB6
mutant enzyme R110A, in 100 mM sodium citrate buffer (pH 5.0), at 30C
0.3
-
cannabigerolate
-, Q8GTB6
wild type enzyme, in 100 mM sodium citrate buffer (pH 5.0), at 30C
0.888
-
cannabigerolate
-
recombinant enzyme, in 100 mM sodium citrate (pH 5.5), at 30C
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.096
-
cannabigerolate
-, Q8GTB6
mutant enzyme R108A, in 100 mM sodium citrate buffer (pH 5.0), at 30C; mutant enzyme R110A, in 100 mM sodium citrate buffer (pH 5.0), at 30C
323294
0.556
-
cannabigerolate
-, Q8GTB6
wild type enzyme, in 100 mM sodium citrate buffer (pH 5.0), at 30C
323294
6.78
-
cannabigerolate
-
recombinant enzyme, in 100 mM sodium citrate (pH 5.5), at 30C
323294
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.078
-
-
unpurified recombinant enzyme, in 100 mM sodium citrate (pH 5.5), at 30C
0.41
-
-
recombinant enzyme after 5250fold purification, in 100 mM sodium citrate (pH 5.5), at 30C
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.4
-
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
the enzyme is localized to and functions in the storage cavity of glandular trichomes
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
74000
-
-
gel filtration
76000
-
-
about 76000 Da, gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
Q8GTB6
x * 58600, calculated from amino acid sequence; x * 60000, SDS-PAGE
?
-
x * 58600, mature enzyme, calculated from amino acid sequence; x * 59000, deglycosylated enzyme, SDS-PAGE; x * 79000, glycosylated enzyme, SDS-PAGE
?
-, Q8GTB6
x * 58597, mature protein, calculate from amino acid sequence; x * 60000, recombinant enzyme, SDS-PAGE; x * 75000, native enzyme, SDS-PAGE
monomer
-
1 * about 75000, SDS-PAGE
monomer
-
1 * 75000, SDS-PAGE
monomer
-
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
Q8GTB6
the enzyme possesses at least eight potential N-linked glycosylation sites
glycoprotein
-
the enzyme contains seven possible Asn glycosylation sites. Endoglycosidase treatment affords a deglycosylated enzyme with more catalytic activity than that of the glycosylated form
glycoprotein
-, Q8GTB6
the enzyme contains eight possible Asn glycosylation sites
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
sitting drop vapor diffusion method, in 0.09 M HEPES buffer pH 7.5 containing 1.26 M sodium citrate
Q8GTB6
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
DE-52 column chromatography, phenyl Sepharose CL-4B column chromatography, and hydroxylapatite column chromatography
-
diethylaminoethyl-cellulose column chromatography, phenyl Sepharose CL-4B column chromatography, and hydroxylapatite column chromatography
-
hydroxylapatite column chromatography
-
Toyopearl CM-650 column chromatography, gel filtration
Q8GTB6
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Nicotiana tabacum BY-2 cells
-
expressed in Sf9 insect cells
-
expressed in the methylotrophic Pichia pastoris strain SMD1168h
-
expressed using a baculovirus insect expression system and expressed in transgenic Nicotiana tabacum hairy roots and methylotrophic yeast Pichia pastoris
-
using a baculovirus-Sf9 insect cell expression system and expressed in Nicotiana tabacum hairy roots
-, Q8GTB6
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the secreted enzyme activity in Pichia pastoris cultures is increased to 472% in the presence of 0.5% (w/v) casamino acids, 277% in the presence of proteinase inhibitors (10 mg/l 4-(2-aminoethyl) benzensulfonyl fluoride, 1 mg/l pepstatin A, and 1 mg/l E-64), 226% in the presence of 10 mg/ml FAD, 248% in the presence of 10 mg/ml FMN, 323% in the presence of 10 mg/ml riboflavin, and 298% in the presence of 1% (v/v) methanol
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
E442Q
-
the enzyme shows 85% of wild type activity
H114A
-, Q8GTB6
the inactive mutant contains no FAD
H114A
-
the mutation completely abolishes enzymatic activity
H208A
-, Q8GTB6
the mutant shows 94% wild type activity
H292A
-
the enzyme shows 5% of wild type activity
H92A
-, Q8GTB6
the mutant shows 87% wild type activity
R108A
-, Q8GTB6
the mutant shows 13% wild type activity
R110A
-, Q8GTB6
the mutant shows 10% wild type activity
Y417F
-
the enzyme shows 42% of wild type activity
Y484F
-
the mutation completely abolishes enzymatic activity