Production of Δ1-tetrahydrocannabinolic acid by the biosynthetic enzyme secreted from transgenic Pichia pastoris

Futoshi Taura, Emi Dono, Supaart Sirikantaramas, Kohji Yoshimura, Yukihiro Shoyama, Satoshi Morimoto

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Δ1-Tetrahydrocannabinolic acid (THCA) synthase is the enzyme that catalyzes the oxidative cyclization of cannabigerolic acid into THCA, the acidic precursor of Δ1-tetrahydrocannabinol. We developed a novel expression system for THCA synthase using a methylotrophic yeast Pichia pastoris as a host. Under optimized conditions, the transgenic P. pastoris secreted ∼1.32 nkat/l of THCA synthase activity, and the culture medium, from which the cells were removed, effectively synthesized THCA from cannabigerolic acid with a ∼98% conversion rate. The secreted THCA synthase was readily purified to homogeneity. Interestingly, endoglycosidase treatment afforded a deglycosylated THCA synthase with more catalytic activity than that of the glycosylated form. The non-glycosylated THCA synthase should be suitable for structure-function studies because it displayed much more activity than the previously reported native enzyme from Cannabis sativa as well as the recombinant enzyme from insect cell cultures.

Original languageEnglish
Pages (from-to)675-680
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume361
Issue number3
DOIs
Publication statusPublished - Sep 28 2007

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Pichia
Acids
Enzymes
Dronabinol
Glycoside Hydrolases
Cyclization
Cannabis
Cell culture
Yeast
Culture Media
Catalyst activity
Insects
Cell Culture Techniques
Yeasts

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Production of Δ1-tetrahydrocannabinolic acid by the biosynthetic enzyme secreted from transgenic Pichia pastoris. / Taura, Futoshi; Dono, Emi; Sirikantaramas, Supaart; Yoshimura, Kohji; Shoyama, Yukihiro; Morimoto, Satoshi.

In: Biochemical and Biophysical Research Communications, Vol. 361, No. 3, 28.09.2007, p. 675-680.

Research output: Contribution to journalArticle

Taura, Futoshi ; Dono, Emi ; Sirikantaramas, Supaart ; Yoshimura, Kohji ; Shoyama, Yukihiro ; Morimoto, Satoshi. / Production of Δ1-tetrahydrocannabinolic acid by the biosynthetic enzyme secreted from transgenic Pichia pastoris. In: Biochemical and Biophysical Research Communications. 2007 ; Vol. 361, No. 3. pp. 675-680.
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