Microbial biosynthesis of rare cannabinoids.

Area of Science:

• Biotechnology
• Synthetic Biology
• Microbial Engineering

Background:

• Cannabis plants produce abundant cannabinoids like ∆9-tetrahydrocannabinol (∆9-THC) and cannabidiol.
• Rare ∆9-THC analogs with unique alkyl chains exhibit potent bioactivity but are difficult to isolate or synthesize.
• Microbial biosynthesis presents a promising alternative for producing these rare cannabinoids.

Purpose of the Study:

• To develop a Saccharomyces cerevisiae host for the microbial biosynthesis of rare cannabinoids.
• To engineer yeast for efficient production of specific rare cannabinoid acids.
• To explore the potential of this platform for generating novel cannabinoids.

Main Methods:

• Engineered Saccharomyces cerevisiae to accumulate geranyl pyrophosphate.
• Overexpressed a fungal pathway for 2,4-dihydroxy-6-alkyl-benzoic acids synthesis.
• Introduced UbiA-prenyltransferase and ∆9-tetrahydrocannabinolic acid synthase genes.

Main Results:

• Successfully biosynthesized two rare cannabinoid acids: C1-substituted ∆9-tetrahydrocannabiorcolic acid (∼16 mg/L) and C7-substituted ∆9-tetrahydrocannabiphorolic acid (∼5 mg/L).
• Thermally decarboxylated ∆9-tetrahydrocannabiphorolic acid to yield ∆9-tetrahydrocannabiphorol.
• Demonstrated the feasibility of producing rare cannabinoids from simple sugars using engineered yeast.

Conclusions:

• Engineered Saccharomyces cerevisiae serves as an effective host for producing rare cannabinoids.
• The microbial platform has potential for producing a diverse range of rare and novel cannabinoids by leveraging fungal gene clusters.
• This approach offers a sustainable and scalable method for accessing valuable cannabinoid compounds.