Fig. 1
Introduction of unnatural amino acid into eGFP and high throughput screening. a The amino acid sequence of Lys-eGFP was extended with two Gly after position 1 and the unnatural amino acid was incorporated at the position of the amber stop codon TAG which was introduced at position 4 using (b) propargyl-L-lysine (Plk, 1) or (S)-2-amino-6-((2-azidoethoxy)carbonylamino) hexanoic acid (Alk, 2). c Beta barrel structure of eGFP (PDB ID: 2Y0G) with highlighted chromophore in the center and incorporation site for the unnatural amino acid at the N-terminus at position 4. d Non-invasive online monitoring of E. coli cultures, producing the unnatural amino acid containing eGFP. The process parameters unnatural amino acid concentration (cuAA), time of unnatural amino acid addition (tuAA), IPTG concentration (cIPTG) and time of IPTG addition (tIPTG) were studied in parallelized experiments