Fig. 3: Extension of the THETA cycle in vitro.
From: Construction and modular implementation of the THETA cycle for synthetic CO2 fixation
a, A simplified scheme of THETA 3.9.9.mc. Modifications from THETA 3.9.9 in a and c are marked in red. ATP and NAD(P)H regeneration systems in a and c are the same as in THETA 3.9.9 and are not shown. b, For the production of malonyl-CoA from CO2, we followed the same protocol used in Fig. 2d and added Pcc* (PccD407I from Methylobacterium extorquens) to carboxylate acetyl-CoA. The cycle was tested with starting concentrations of 200 µM and 50 µM pyruvate, which had provided the highest yield and the highest turnover rates, respectively, for THETA version 3.9.9 (Fig. 2d). c, A simplified scheme of THETA 3.9.9.gc. d, For the production of glycolate, we replaced part of the reaction sequence as shown in c and used 50 µM and 200 µM succinate to start the reaction sequence. Because the release of the primary CO2-fixation product (glyoxylate) occurs at the level of the Icl reaction, we used succinate as starting substrate (instead of pyruvate). Data in b and d represent the mean ± s.d. obtained in n = 3 independent experiments. Pcc, propionyl-CoA carboxylase; Cs, citrate synthase; Acn, aconitase; Icl, isocitrate lyase; Gor, glyoxylate reductase.
