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Production of D1-tetrahydrocannabinolic acid by the biosynthetic enzyme secreted from transgenic Pic

Production of D1-tetrahydrocannabinolic acid by the biosynthetic enzyme secreted from transgenic Pic

  1. jwh210
    D1-Tetrahydrocannabinolic acid (THCA) synthase is the enzyme that catalyzes the oxidative cyclization of cannabigerolic acid into
    THCA, the acidic precursor of D1-tetrahydrocannabinol. We developed a novel expression system for THCA synthase using a methy-
    lotrophic 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 affor-
    ded a deglycosylated THCA synthase with more catalytic activity than that of the glycosylated form. The non-glycosylated THCA syn-
    thase 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.