Production of Delta(1)-tetrahydrocannabinolic acid by the biosynthetic enzyme secreted from transgenic Pichia pastoris.
Futoshi Taura, Emi Dono, Supaart Sirikantaramas, Kohji Yoshimura, Yukihiro Shoyama, Satoshi Morimoto
Biochemical and Biophysical Research Communications September 28, 2007 DOI: 10.1016/j.bbrc.2007.07.079 (opens in new tab)
Study at a glance
AI-extracted from the abstract| Characteristics | Laboratory experimental study (recombinant enzyme expression and characterization) Peer reviewed |
|---|---|
| Population | THCA synthase expressed in transgenic Pichia pastoris yeast cultures |
| Topics | Cannabis |
| Key findings | Transgenic Pichia pastoris secreted about 1.32 nkat/l of THCA synthase activity, and the cell-free culture medium converted cannabigerolic acid to THCA at approximately 98%. Deglycosylated THCA synthase showed more catalytic activity than the glycosylated form and, according to the authors, more activity than the native Cannabis sativa enzyme and a recombinant enzyme from insect cell cultures, making it suitable for structure-function studies. |
Abstract
Delta(1)-Tetrahydrocannabinolic acid (THCA) synthase is the enzyme that catalyzes the oxidative cyclization of cannabigerolic acid into THCA, the acidic precursor of Delta(1)-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 approximately 1.32nkat/l of THCA synthase activity, and the culture medium, from which the cells were removed, effectively synthesized THCA from cannabigerolic acid with a approximately 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.
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