Fig. 5: Screening a THCAS mutant yeast library for high producers.
From: Screening microbially produced Δ9-tetrahydrocannabinol using a yeast biosensor workflow
a Workflow for screening THC from microbial production using the yeast CB2 biosensor. AHF, acetonitrile/H2O/formic acid (80/20/0.05 %). b Screening a library of 108 THCAS mutant yeast strains for increased THCA production using the CB2 biosensor, highlighting five stains with increased THC titres over the unmutated THCAS (black). Experimental measurements are GFP levels per cell as determined on a plate reader and shown as the mean ± SD from two biosensor measurements of a single extracted sample. Measurements were normalised to no ligand (0, dotted line) and unmutated THCAS control (1, solid line). Samples with StDev > 5 % of the maximum signal were omitted (1/109 samples). c Relative quantification of THC from the 108 samples shown in b by LC-MS, highlighting the five strains identified as high producers using the CB2 biosensor (black). Experimental measurements are relative THC amounts (mAU) as determined by LC-MS and shown as individual values. d Correlation of the CB2 biosensor response with relative quantification of THC by LC-MS, demonstrating a Pearson’s correlation coefficient of 0.6, p (two-tailed) < 0.0001. Data are mean values from b and individual values from c for the 108 decarboxylated cell extracts. A straight-line curve was fitted using GraphPad Prism linear regression fit, R2 = 0.36.
