Screening microbially produced Δ9-tetrahydrocannabinol using a yeast biosensor workflow
W. M. Shaw, Yunfeng Zhang, Xinyu Lu, Ahmad S. Khalil, Graham Ladds, Xiaozhou Luo, Tom Ellis
Abstract
Abstract Microbial production of cannabinoids promises to provide a consistent, cheaper, and more sustainable supply of these important therapeutic molecules. However, scaling production to compete with traditional plant-based sources is challenging. Our ability to make strain variants greatly exceeds our capacity to screen and identify high producers, creating a bottleneck in metabolic engineering efforts. Here, we present a yeast-based biosensor for detecting microbially produced Δ 9 -tetrahydrocannabinol (THC) to increase throughput and lower the cost of screening. We port five human cannabinoid G protein-coupled receptors (GPCRs) into yeast, showing the cannabinoid type 2 receptor, CB2R, can couple to the yeast pheromone response pathway and report on the concentration of a variety of cannabinoids over a wide dynamic and operational range. We demonstrate that our cannabinoid biosensor can detect THC from microbial cell culture and use this as a tool for measuring relative production yields from a library of Δ 9 -tetrahydrocannabinol acid synthase (THCAS) mutants.