Engineering cannabinoid production in Saccharomyces cerevisiae

Area of Science:

• Biotechnology
• Synthetic Biology
• Metabolic Engineering

Background:

• Phytocannabinoids possess valuable pharmacological properties but plant extraction faces limitations.
• Current heterologous production systems for cannabinoids lack industrially viable titers.
• Developing alternative production methods is crucial for meeting demand.

Purpose of the Study:

• To establish a robust yeast system for producing cannabinoids from simple feedstocks.
• To enhance cannabinoid production titers through protein engineering and metabolic pathway optimization.
• To demonstrate the de novo biosynthesis of specific cannabinoids in a heterologous host.

Main Methods:

• Engineered a yeast system utilizing the aromatic prenyltransferase NphB.
• Optimized precursor (olivetolic acid) production to 56 mg/L.
• Implemented cannabinoid synthase genes and utilized a fed-batch approach for enhanced production.

Main Results:

• Achieved production of cannabigerolic acid and cannabigerol from glucose and hexanoic acid.
• Significantly increased production via an ERG20WW-NphB fusion protein.
• Demonstrated first-time heterologous biosynthesis of cannabichromenic acid in yeast.
• Showcased pH-dependent product spectrum of vacuole-localized cannabinoid synthases.
• Attained titers of 18.2 mg/L for cannabigerolic acid and 117 mg/L for olivetolic acid.

Conclusions:

• Developed a highly efficient heterologous yeast platform for cannabinoid biosynthesis.
• The engineered system offers a scalable and controllable alternative to plant extraction.
• Future research can leverage this platform for producing a wider range of cannabinoids.