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. 2022 Mar 8;11(3):1114–1128. doi: 10.1021/acssynbio.1c00342

Figure 2.

Figure 2

Figure 2

Lyoprotectant effects of five sugars individually added to the two cell-free formulations. CFPS based on PEP (A–E) and MDX (F–J) and dehydrated either by high-cost lyophilization or by the low-cost drying method. Samples were dried and stored at room temperature for 1 day and 2 weeks. Cell-free reactions were rehydrated and incubated at 29 °C for 15 h. The final concentrations of additives in the reactions are indicated on the horizontal axes. The percentage of the recovered protein production was calculated relative to that seen in fresh, additive-free reactions with the energy sources PEP or MDX. Error bars represent standard error over three technical measurements. (K) Effects of lactose on protein yields in fresh cell-free reactions based on PEP or MDX as energy sources. Samples were supplemented with lactose, 11.2 mM (PEP mixture; Tables S2 and S5) and 13.7 mM (MDX mixture; Tables S1 and S5) as indicated. Cell-free reactions for the production of a green fluorescent protein were incubated at 29 °C for 15 h using psfGFP as a DNA template. Yields were calculated relative to fluorescence values seen in PEP-formulated cell-free reactions in the absence of lactose. Error bars represent standard error over three technical measurements.