Fig. 2

Quantification of miltiradiene and abietatriene in engineered C. glutamicum. (A) GC-MS measurement of miltiradiene (C₂₀H₃₂) and abietatriene (C₂₀H₃₀) produced in engineered C. glutamicum. 1-eicosene (C₂₀H₄₀) was injected into sample as internal standard and ferruginol was used for quantification of miltiradiene and abietatriene (ferruginol equivalent mg/gDCW). (B) Heterologous diterpenes (miltiradiene and abietatriene) were produced in engineered C. glutamicum strains harboring various plasmids. A two-plasmid system was used for gene expressions: a pBL1-origin vector harboring GGPPS and OverMEP modules and a pCG1-origin vector harboring DiTPS module. Miltiradiene (grey) and abietatriene (white) were quantified as ferruginol equivalent mg/gDCW using GC-FID. Data represent mean values of at least triplicated cultivations, and error bars represent standard deviations. LK stands for a linker