Fig. 4. Cell-free reactions with extract from engineered yeast strains enable production of diverse metabolites.
a Metabolic map showing the conversion of glucose to glycerol via dihydroxyacetone phosphate (DHAP) and glycerol-3-phosphate (G3P) or itaconic acid via citrate and cis-aconitate (c-A). b, c Initial cell-free reaction conditions confirm that strains engineered for increased product titers result in cell extracts with greater flux toward products of interest (***p ≤ 0.001 compared to base strain). From a two-tailed Student’s t-test, p = 8.45 × 10−5 for itaconic acid, p = 7.16 × 10−6 for glycerol, and p = 2 × 10−3 for ethanol. d-e Yeast cell-free reactions with optimized conditions rapidly convert glucose into products, resulting in comparable concentrations to in vivo efforts in significantly less time. Data represent mean ± standard deviation of n = 3 technical replicates, and time courses represent separate samples quenched at each time point. Source data are provided as a Source Data file.