Biosynthesis of cannabinoid precursor olivetolic acid in genetically engineered Yarrowia lipolytica

Date

2022-11-12

Department

Program

Citation of Original Publication

Ma, J., Gu, Y. & Xu, P. Biosynthesis of cannabinoid precursor olivetolic acid in genetically engineered Yarrowia lipolytica. Commun Biol 5, 1239 (2022). https://doi.org/10.1038/s42003-022-04202-1

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Attribution 4.0 International (CC BY 4.0)

Subjects

Abstract

Engineering microbes to produce plant-derived natural products provides an alternate solution to obtain bioactive products. Here we report a systematic approach to sequentially identify the rate-limiting steps and improve the biosynthesis of the cannabinoid precursor olivetolic acid (OLA) in Yarrowia lipolytica. We find that Pseudomonas sp LvaE encoding a short-chain acyl-CoA synthetase can efficiently convert hexanoic acid to hexanoyl-CoA. The co-expression of the acetyl-CoA carboxylase, the pyruvate dehydrogenase bypass, the NADPH-generating malic enzyme, as well as the activation of peroxisomal β-oxidation pathway and ATP export pathway are effective strategies to redirect carbon flux toward OLA synthesis. Implementation of these strategies led to an 83-fold increase in OLA titer, reaching 9.18 mg/L of OLA in shake flask culture. This work may serve as a baseline for engineering cannabinoids biosynthesis in oleaginous yeast species.