Engineered yeast unlocks sustainable production of cannabinoids and analogs

By reprogramming yeast metabolism, the team achieved de novo biosynthesis of medically valuable cannabinoids, laying the foundation for scalable, climate-independent production methods.

The endocannabinoid system regulates fundamental functions such as memory, stress response, appetite, and immune signaling. Among its plant-derived modulators, cannabidiol (CBD) has received clinical recognition, with the U.S. Food and Drug Administration approving Epidiolex® for certain seizure disorders. Other cannabinoids, such as cannabigerolic acid (CBGA), have shown promise in preclinical studies as anticonvulsants, metabolic regulators, and even antiviral agents with activity against SARS-CoV-2. Yet access to these compounds remains limited by agricultural yields, climate variability, and extraction costs. Microbial engineering provides an alternative, offering controlled, climate-independent production that can be scaled to industrial levels. While the yeast Saccharomyces cerevisiae has been used to produce cannabinoids at moderate yields, Y. lipolytica offers distinct advantages: it generates high acetyl-CoA and malonyl-CoA flux, grows on low-cost substrates, and has expanded membrane space to support complex enzymatic activity. Classified as “generally regarded as safe” (GRAS), this oleaginous yeast represents a promising platform for cannabinoid biosynthesis.

A study (DOI:10.1016/j.bidere.2025.100021) published in BioDesign Research on 3 April 2025 by Peng Xu, Gil Bar-Sela & Idan Cohen’s team, Guangdong Technion-Israel Institute of Technology & Technion-Israel Institute of Technology, demonstrates the potential of Yarrowia lipolytica as a sustainable microbial platform for scalable production of diverse cannabinoids and their analogs, offering new opportunities for pharmaceutical development and biotechnology.

The research team employed a stepwise metabolic engineering strategy to reprogram Yarrowia lipolytica for cannabinoid biosynthesis, focusing on systematically removing metabolic bottlenecks and enhancing precursor supply. To improve the polyketide module responsible for olivetolic acid (OLA) synthesis, they selected the PpLvaE enzyme from Pseudomonas putida to efficiently convert hexanoic acid into hexanoyl-CoA, integrated multiple OLA gene cassettes at different chromosomal loci, and optimized feeding strategies to reduce hexanoic acid toxicity. Redirecting carbon flux through DGA1 knockout and ylACC1 overexpression further boosted malonyl-CoA availability, raising OLA production to 6.73 mg/L. In parallel, the isoprenoid pathway was enhanced by introducing MVA-pathway boosters (EfmvaE, EfmvaS) and geranyl pyrophosphate (GPP)-biased ERG20 mutants from Y. lipolytica and S. cerevisiae. To overcome prenyltransferase (PTase) limitations, the team amplified copy numbers of truncated CsPT4, evaluated subcellular localizations, and co-expressed NphB (Y288A/G286S) to create condensate-like dual PTase assemblies. These strategies increased cannabigerolic acid (CBGA) titers from sub-milligram levels to 2.85 mg/L, and further supplementation with OLA plus ERG7 inhibition elevated production to 15.7 mg/L—the highest reported in Y. lipolytica. Expanding beyond CBD-C5 precursors, the researchers introduced the noncanonical polyketide synthase ArmB from Armillaria mellea, enabling de novo orsellinic acid (OSA) biosynthesis at up to 18.87 mg/L under fed-batch cultivation. This pathway provided the basis for cannabigerorcinic acid (CBGOA), a CBD-C1 analog precursor, which reached 541 μg/L with dual PTase expression. Overall, by integrating precursor optimization, enzyme co-assembly, and pathway expansion, the team unlocked de novo microbial production of OLA, CBGA, OSA, and CBGOA, demonstrating Y. lipolytica’s versatility as a chassis for sustainable and scalable cannabinoid biosynthesis.

This work marks the first successful de novo production of CBGA, CBGOA, and OSA in Y. lipolytica. By combining pathway optimization, novel enzyme integration, and biomolecular condensate-like strategies, researchers have established a foundation for sustainable cannabinoid biosynthesis. The study not only advances cannabinoid research but also sets the stage for broader applications of engineered Y. lipolytica in biotechnology and medicine.

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References

DOI

10.1016/j.bidere.2025.100021

Original Source URL

https://doi.org/10.1016/j.bidere.2025.100021

Funding information

The funding is supported by the National Natural Science Foundation of China (general grant 22378083) and the Li Ka-shing Foundation (EN2400022) and the Muyuan Laboratory (Program ID: 12106022401).

About BioDesign Research

BioDesign Research is dedicated to information exchange in the interdisciplinary field of biosystems design. Its unique mission is to pave the way towards the predictable de novo design and assessment of engineered or reengineered living organisms using rational or automated methods to address global challenges in health, agriculture, and the environment.