Fig. 4

Generating the strains for continuous bioproduction of linalool. To generate the parental strain, the DH5α strain was transformed with pMVA-Lin. Terpenoid synthesis can occur either through the native 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway via dxr, or through the heterologous mevalonate (MVA) pathway with IPTG induction. To generate the Δdxr strain, DH5α dxrFRT was first transformed with pTet-Flp and pMVA-Lin. Anhydrotetracycline (aTc) induced the expression of the Flp recombinase from pTet-Flp, which knocked out the dxr gene. Terpenoid synthesis was restored by inducing the heterologous MVA pathway on pMVA-Lin with IPTG. The Δdxr strain was obtained by isolating pure colonies with a verified knockout. This strain relies only on the said MVA pathway for terpenoid synthesis