Fig. 3. Prototyping and optimization of feedback modules 4a–d.

Methylmalonyl-CoA formation from 250 µM glyoxylate through anaplerotic feedback modules 4a–d of varying compositions is presented. All experiments contained modules 1, 2 and 3 together with one of the four anaplerotic modules, in which selected enzymes and/or cofactors were varied. a, Module 4a using different amounts of Frd (10, 50 and 100 µU ml⁻¹ = 0.01 to 0.1 mU ml⁻¹). All experiments contained 50 mM formate, except ++form, which contained 100 mM formate. b, Module 4b with different amounts of Mdh. c, Module 4c with or without adenylate kinase (Adk). d, Module 4d with different amounts of CoA. All experiments were performed in technical triplicates and are displayed as mean ± s.d.

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