Table 2.

Chart of fermentation conditions and protein yields. Host refers to the host E. coli strain for fermentation.

Setup	Model protein	Host[a]	Met source[b]	Azide[c]	PA[d]	Ind. time[e]	Protein yield[f] [mg/L]	Label yield[g]
1	B*	M	0.06 mM Met, 8 h	0.8 mM, 1 h	–	o/n	1.5	1/2
2	GTL	M	0.06 mM Met, 8 h	0.8 mM, 1 h	–	o/n	5.0	6/7
3	GTL	M	0.06 mM Met, o/n	0.8 mM, 3 h	–	5 h	1.5	5/7
4	ECFP‐N	A	0.06 mM Met, 7 h	0.8 mM, 1 h	–	o/n	14.1	3/6
5	ECFP‐C	A	0.06 mM Met, 6 h	0.8 mM, 3 h	–	o/n	2.6	3/5
6	GFP‐1M	A	0.06 mM Met, 7 h	0.8 mM, 1 h	–	o/n	2.7	0/1
7	GFP‐2M	A	0.06 mM Met, 7 h	0.8 mM, 1 h	–	o/n	5.7	1/2
8	ECFP‐N	A	3.5 g/L YE, 8 h	2.4 mM, 4.5 h [h]	1 mM	20 h	36.0	5/6
9	GFP‐1M	A	3.5 g/L YE, 8 h	2.4 mM, 4.5 h [h]	1 mM	20 h	9.7	1/1
10	GFP‐2M	A	3.5 g/L YE, 8 h	2.4 mM, 4.5 h [h]	1 mM	20 h	20.0	2/2
11	ECFP‐N	A	3.5 g/L YE, 8 h	5.6 mM, 4.5 h[i]	–	20 h	14.0	1/6

[a] M: MDS15 and A: MDS15A; [b] The methionine source and its feeding time; [c] The feeding strategy of sodium azide; [d] Addition of pantothenic acid. [e] Length of induction time. [f] Amount of purified protein, expressed in mg per L culture. [g] Detected Aha residues divided by total Met positions in the target proteins, as determined by MS analysis (In the table, yield results should be taken as a rough estimate; a more thoroughly analysis of the labeling yield was performed by dOG conjugation and is discussed in the text). Protein and label yield values are the average of at least three determinations. The range of values was always within ±5 % from the average value. [h] For setups 8–10, supplying of 0.8 mM sodium azide was performed 3 times every 1.5 h. [i] For setup 11, 0.8 mM sodium azide was added 5 times. in bold, the most successful setups (described in the text).