(A) Kinetic linearity of overall reconstituted pathway. Reaction conditions: 2–10 μM AccAD, AccC, 6–30 μM holo-AccB, 0.2–1 μM FabABDFGH, 2–10 μM FabIZ, 6–30 μM holo-ACP, 6–30 μM TesA, 600 μM acetyl-CoA, 15 mM bicarbonate, 1 mM ATP, 5 mM MgCl2, and 1.3 mM NADPH. (B) Palmitic acid product formation in 5-fold dilution system. (C) Initial reaction rates of optimized reconstituted systems with an individual enzyme concentration doubled. Experiments were performed in two-fold dilution system.