Development of an efficient yeast platform for cannabigerolic acid biosynthesis

Area of Science:

• Biotechnology
• Metabolic Engineering
• Synthetic Biology

Background:

• Cannabinoids, like cannabigerolic acid (CBGA), possess significant therapeutic potential for various human ailments, including cancer and SARS-CoV-2 infections.
• Cannabis sativa prenyltransferase (CsPT4) synthesizes CBGA, but microbial production in Saccharomyces cerevisiae yields suboptimal titers due to metabolic bottlenecks and enzyme limitations.

Purpose of the Study:

• To engineer Saccharomyces cerevisiae for enhanced production of cannabigerolic acid (CBGA).
• To overcome limitations in hexanoate conversion and improve the activity and stability of the Cannabis sativa prenyltransferase (CsPT4) enzyme.

Main Methods:

• Metabolic engineering strategies were employed to reduce hexanoate consumption via the beta-oxidation pathway and minimize its incorporation into fatty acids.
• Cellular engineering involved expanding the endoplasmic reticulum and fusing an auxiliary protein to CsPT4 to enhance its performance.
• The engineered yeast chassis was cultivated using glucose and hexanoate as substrates.

Main Results:

• The engineered Saccharomyces cerevisiae exhibited a significant 78.64-fold increase in CBGA production.
• The final CBGA titer reached 510.32 ± 10.70 mg L⁻¹.
• The combined metabolic and protein engineering strategies effectively addressed the challenges in microbial CBGA synthesis.

Conclusions:

• The developed engineered yeast strain represents a substantial advancement in microbial CBGA production.
• This enhanced production platform holds promise for the cost-effective and scalable synthesis of therapeutic cannabinoids.
• Further optimization could lead to even higher yields for pharmaceutical applications.