Fig. 4: Solubility and in vitro productivity of high solubility library variants.
From: Biosensor Guided Polyketide Synthases Engineering for Optimization of Domain Exchange Boundaries
a Solubility measurement using ΔarsB::Pibp GFP biosensor strain of 40 high solubility colonies, a subset of which have their junction positions marked and were selected for (b) SDS-PAGE quantification of PKS abundance in different protein fractions. (c) In vitro reaction scheme with the synthetic starter 1 and methyl- or malonyl-CoA as substrate. DESBM6 AT natively accepts methylmalonyl-CoA while EpoM4 AT can accept both methyl- and malonyl-CoA. In vitro production of methyl TKL after 1 and 24 hours (d) and desmethyl TKL after 24 hours (e). DEBSM6 is the parental PKS, D1 is included as a reference to what is currently known as the optimal junctions for domain exchange. D0 was excluded due to it already been shown to be inactive8. All strains were induced with 250 µM IPTG. Data is presented as mean values of three biological replicates, dots are individual data points. Arb. units = arbitrary units. Source data are provided as a Source Data file.
