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. 2021 Aug 31;7(9):1500–1507. doi: 10.1021/acscentsci.1c00499

Figure 3.

Figure 3

Yield and purity of sfGFP containing a single ncAA when expressed in Vmax X2 versus E. coli strains. (A) Isolated yield (mg/L) and (B) fidelity (%) of sfGFP containing the indicated ncAA at position 2 when expressed in the indicated strain. Cells were grown for 4 h and lysed via sonication, and sfGFP was isolated using IMAC. Yields were determined using the Pierce 660 nm protein assay (Thermo Scientific) and a BSA standard curve. Fidelity was determined from LC-MS/MS data as the fraction of the Glu-C-generated N-terminal peptide MXKGEE containing the desired ncAA (X) at position 2 relative to all other detectable amino acids at that position. The dashed line indicates 100% fidelity. Additional LC-MS/MS data are found in Figures S4 and S5.