Table 1.

Proteome-wide [3,2]Tpa incorporation followed by mass spectrometric and amino acid analyses. The MT-strains were isolated at different time points throughout the long term evolution experiment (Figure 2) and represent increasing stages of adaptation. Please note that [3,2]Tpa is the amino acid version of the Trp analog, which is produced introcellularily by the strain. In contrast, [3,2]Tp is the Ind analog version which is supplied to the medium. An extended version of this table with additional intermediates can be found in the SI (Table S5).

Strain[a]	MT0	MT0	MT1	MT20
MS/MS[b]
Trp [%]	99.5 ±1.2	99.7 ± 0.5	0.6 ± 0.8	0.5 ± 0.8
[3,2]Tpa [%]	0.5 ±1.2	0.6 ± 0.5	99.4 ± 0.8	99.5 ± 0.8
Amino acid anaylsis[c]
Trp [%]	100 ± 3.8	98.7 ± 3.2	1.6 ± 0.1	-[d]
Medium
Aa concent[e]	19	19	19	0
Ind [μM]	100	100	0	0
[3,2]Tp [μM]	0	25	25	100

^[a]Strain details are provided in section 4 and 6 in the SI;

^[b]Measured peptide occurrence of [3,2]Tpa and Trp measured by MS/MS. 20 of the most abundant tryptophan containing peptides were chosen for quantification;

^[c]GC/MS analysis of hydrolysed proteome samples derivatized by esterification and amidiation.

^[d]no peak observed in the chromatogram;

^[e]Amino acid content of the medium. A detailed overview of the medium composition can be found in section 4 in the SI.