Fig. 4. Prototyping and application of the different anaplerotic pathways for 6-DEB production.
a, Methylmalonyl-CoA formation of the different anaplerotic modules 4a, 4b, 4c, or 4d when coupled to modules 1–3. Experiments were started with 250 µM glyoxylate (coupled modules 1, 2, 3 plus either module 4a, 4b, 4c or 4d) or 167 µM propionyl-CoA (module 1 alone). b, 6-DEB formation by the different anaplerotic pathways modules 4a, 4b, 4c or 4d when coupled to modules 1–3 and DEBS (module 5). Shown also is the 6-DEB formation of module 1 coupled to module 5 without any anaplerotic modules, and module 5 alone, started with propionyl-CoA and methylmalonyl-CoA. Assays were started using either 167 or 333 µM propionyl-CoA. For details and additional data, see Extended Data Fig. 5. *, no 6-DEB detected. **, started with 0.8 mM propionyl-CoA and 1.0 mM methylmalonyl-CoA. All experiments were performed in technical duplicates and are displayed as mean (white squares) of the individual measurements (colored squares).