First direct evidence for the mechanism of .DELTA.1-tetrahydrocannabinolic acid biosynthesis

Taura, Futoshi; Morimoto, Satoshi; Shoyama, Yukihiro; Mechoulam, Raphael

doi:10.1021/ja00143a024

Cited by 172 publications

(167 citation statements)

References 0 publications

Supporting

Mentioning

157

Contrasting

Unclassified

Order By: Relevance

“…THCA synthase oxidatively cyclizes the monoterpene moiety of CBGA to form THCA (9). Similar reactions are often found in monoterpene biosynthesis, where biosynthetic enzymes (monoterpene cyclases) cyclize geranyl pyrophosphate into various monoterpenes (12), but these reactions are not accompanied by oxidation, contrary to the THCA synthase reaction.…”

mentioning

confidence: 65%

“…Similar reactions are often found in monoterpene biosynthesis, where biosynthetic enzymes (monoterpene cyclases) cyclize geranyl pyrophosphate into various monoterpenes (12), but these reactions are not accompanied by oxidation, contrary to the THCA synthase reaction. Because of the novel catalytic property, it is of interest to know the primary structure of THCA synthase, although we revealed no structural information on this enzyme except for the N-terminal sequence containing 15 amino acid residues (9). Moreover, we have not unequivocally determined which subclass this oxidoreductase belongs to, because the coenzyme or cofactor required for the oxidocyclization of CBGA was not identified (9).…”

mentioning

confidence: 87%

“…Because of the novel catalytic property, it is of interest to know the primary structure of THCA synthase, although we revealed no structural information on this enzyme except for the N-terminal sequence containing 15 amino acid residues (9). Moreover, we have not unequivocally determined which subclass this oxidoreductase belongs to, because the coenzyme or cofactor required for the oxidocyclization of CBGA was not identified (9). Thus, the mechanism of the THCA synthase reaction has remained largely unclear.…”

mentioning

confidence: 90%

“…1). Our previous studies on the biosynthetic pathway of cannabinoids demonstrated that THCA, which had been believed to be formed through isomerization of cannabidiolic acid, was actually biosynthesized from cannabigerolic acid (CBGA) (9,10) (Fig. 1).…”

mentioning

confidence: 99%

“…1). Furthermore, we identified an oxidoreductase named THCA synthase, which catalyzes THCA biosynthesis in rapidly expanding leaves of C. sativa (Mexican strain) (9). The activity of this enzyme was not detected in a nonpsychoactive Cannabis strain (CBDA strain) (10,11), which contains cannabidiolic acid as a major cannabinoid and only a trace amount of THCA, indicating that THCA synthase is an important enzyme controlling the psychoactivity of C. sativa.…”

mentioning

confidence: 99%

See 4 more Smart Citations

The Gene Controlling Marijuana Psychoactivity

Sirikantaramas

Morimoto

Ishikawa

et al. 2004

Journal of Biological Chemistry

Self Cite

243

155

View full text Add to dashboard Cite

⌬ 1 -Tetrahydrocannabinolic acid (THCA) synthase is the enzyme that catalyzes oxidative cyclization of cannabigerolic acid into THCA, the precursor of ⌬ 1 -tetrahydrocannabinol. We cloned a novel cDNA (GenBank TM accession number AB057805) encoding THCA synthase by reverse transcription and polymerase chain reactions from rapidly expanding leaves of Cannabis sativa. This gene consists of a 1635-nucleotide open reading frame, encoding a 545-amino acid polypeptide of which the first 28 amino acid residues constitute the signal peptide. The predicted molecular weight of the 517-amino acid mature polypeptide is 58,597 Da. Interestingly, the deduced amino acid sequence exhibited high homology to berberine bridge enzyme from Eschscholtzia californica, which is involved in alkaloid biosynthesis. The liquid culture of transgenic tobacco hairy roots harboring the cDNA produced THCA upon feeding of cannabigerolic acid, demonstrating unequivocally that this gene encodes an active THCA synthase. Overexpression of the recombinant THCA synthase was achieved using a baculovirus-insect expression system. The purified recombinant enzyme contained covalently attached FAD cofactor at a molar ratio of FAD to protein of 1:1. The mutant enzyme constructed by changing His-114 of the wild-type enzyme to Ala-114 exhibited neither absorption characteristics of flavoproteins nor THCA synthase activity. Thus, we concluded that the FAD binding residue is His-114 and that the THCA synthase reaction is FAD-dependent. This is the first report on molecular characterization of an enzyme specific to cannabinoid biosynthesis.

show abstract

mentioning

confidence: 65%

mentioning

confidence: 87%

mentioning

confidence: 90%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

The Gene Controlling Marijuana Psychoactivity

Sirikantaramas

Morimoto

Ishikawa

et al. 2004

Journal of Biological Chemistry

Self Cite

243

155

View full text Add to dashboard Cite

show abstract

Terpenoid‐Biosynthese abseits bekannter Wege: unkonventionelle Cyclasen und ihre Bedeutung für die biomimetische Synthese

2014

View full text Add to dashboard Cite

Cyclisierungsreaktionen von Terpenen und Terpenoiden zählen zu den komplexesten chemischen Reaktionen in der Natur und tragen maßgeblich zur enormen Strukturvielfalt dieser größten Naturstofffamilie bei. Zahlreiche Studien auf chemischer, genetischer und biochemischer Ebene wurden durchgeführt, um mechanistische Einblicke in diese faszinierenden Reaktionen zu erlangen, die von Terpen‐ und Terpenoid‐Cyclasen katalysiert werden. Eine Vielzahl dieser Enzyme konnte seither charakterisiert werden. Nach klassischer Lehrbuchmeinung werden Terpen‐/Terpenoid‐Cyclasen gemäß ihrer Struktur und ihrem Reaktionsmechanismus in zwei große Klassen eingeteilt. Jüngste Entdeckungen neuartiger Terpenoid‐Cyclasen zeigen jedoch, dass das natürliche enzymatische Repertoire deutlich vielseitiger ist, als zunächst gedacht. Dieser Aufsatz stellt eben diese Terpenoid‐Cyclasen in den Fokus, die aus dem Rahmen fallen.

show abstract

Total Synthesis Establishes the Biosynthetic Pathway to the Naphterpin and Marinone Natural Products

et al. 2018

View full text Add to dashboard Cite

The naphterpins and marinones are naphthoquinone meroterpenoids with an unusual aromatic oxidation pattern that is biosynthesized from 1,3,6,8‐tetrahydroxynaphthalene (THN). We propose that cryptic halogenation of THN derivatives by vanadium‐dependent chloroperoxidase (VCPO) enzymes is key to this biosynthetic pathway, despite the absence of chlorine in these natural products. This speculation inspired a total synthesis to mimic the naphterpin/marinone biosynthetic pathway. In validation of this biogenetic hypothesis, two VCPOs were discovered that interconvert several of the proposed biosynthetic intermediates.

show abstract

First direct evidence for the mechanism of .DELTA.1-tetrahydrocannabinolic acid biosynthesis

Cited by 172 publications

References 0 publications

The Gene Controlling Marijuana Psychoactivity

The Gene Controlling Marijuana Psychoactivity

Terpenoid‐Biosynthese abseits bekannter Wege: unkonventionelle Cyclasen und ihre Bedeutung für die biomimetische Synthese

Total Synthesis Establishes the Biosynthetic Pathway to the Naphterpin and Marinone Natural Products

Contact Info

Product

Resources

About