Fig. 2: Combinatorial screen of r-BOX enzyme homologs. | Nature Communications

Fig. 2: Combinatorial screen of r-BOX enzyme homologs.

From: Cell-free prototyping enables implementation of optimized reverse β-oxidation pathways in heterotrophic and autotrophic bacteria

Fig. 2

All r-BOX enzymes were used at 0.3 μM with 0.15 μM of Ec_tesA and incubated at 30 °C for 24 h. n = 3, error bars = not shown (see Source Data file for complete data and enzyme abbreviations). The enzymes kept constant during the assay are Ck_thlA, Ck_hbd1, Ca_crt, Td_ter and Ec_tesA if they are not the ones investigated in the panel. A TL × HBD. B HBD × CRT. C CRT × TER. D TER × TL. Full results in Supplementary sheet 1. E Time-course SAMDI-CoA experiment showing dynamics of r-BOX assays over time. Maximum build-up of decanoyl-CoA occurs at 10 h using Re_bktB as the thiolase. All conditions were run and measured in quadruplicate and the average is displayed. F GC-MS characterization of the time course conditions with the addition of 0.15 μM of either Ec_tesA or Ec_fadM termination enzyme. Data represent n = 3 independent experiments, with the standard deviation and mean shown.

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