Literature summary for 1.21.3.7 extracted from

Taura, F.; Dono, E.; Sirikantaramas, S.; Yoshimura, K.; Shoyama, Y.; Morimoto, S.
Production of DELTA1-tetrahydrocannabinolic acid by the biosynthetic enzyme secreted from transgenic Pichia pastoris (2007), Biochem. Biophys. Res. Commun., 361, 675-680.

Search for more literature for 1.21.3.7

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in the methylotrophic Pichia pastoris strain SMD1168h	Cannabis sativa

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.131	-	cannabigerolate	recombinant enzyme, in 100 mM sodium citrate (pH 5.5), at 30°C	Cannabis sativa

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
58600	-	x * 58600, mature enzyme, calculated from amino acid sequence	Cannabis sativa
59000	-	x * 59000, deglycosylated enzyme, SDS-PAGE	Cannabis sativa
79000	-	x * 79000, glycosylated enzyme, SDS-PAGE	Cannabis sativa

Organism

Organism	UniProt	Comment	Textmining
Cannabis sativa	-	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
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	Cannabis sativa

Purification (Commentary)

Purification (Comment)	Organism
hydroxylapatite column chromatography	Cannabis sativa

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.078	-	unpurified recombinant enzyme, in 100 mM sodium citrate (pH 5.5), at 30°C	Cannabis sativa
0.41	-	recombinant enzyme after 5250fold purification, in 100 mM sodium citrate (pH 5.5), at 30°C	Cannabis sativa

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
cannabigerolate + O2	about 98% conversion rate in 24 h	Cannabis sativa	DELTA9-tetrahydrocannabinolate + H2O2	-	?

Subunits

Subunits	Comment	Organism
?	x * 58600, mature enzyme, calculated from amino acid sequence	Cannabis sativa
?	x * 59000, deglycosylated enzyme, SDS-PAGE	Cannabis sativa
?	x * 79000, glycosylated enzyme, SDS-PAGE	Cannabis sativa

Synonyms

Synonyms	Comment	Organism
DELTA1-tetrahydrocannabinolic acid synthase	-	Cannabis sativa
THCA synthase	-	Cannabis sativa

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.888	-	cannabigerolate	recombinant enzyme, in 100 mM sodium citrate (pH 5.5), at 30°C	Cannabis sativa

Expression

Organism	Comment	Expression
Cannabis sativa	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	up

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
6.78	-	cannabigerolate	recombinant enzyme, in 100 mM sodium citrate (pH 5.5), at 30°C	Cannabis sativa

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Release 2023.1 (January 2023)