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Ehrlich, Michael; Gattner, Michael J.; Viverge, Bastien; Bretzler, Johanna; Eisen, David; Stadlmeier, Michael ORCID: 0000-0001-7806-3077; Vrabel, Milan; Carell, Thomas ORCID: 0000-0001-7898-2831 (2. April 2015): Orchestrating the biosynthesis of an unnatural pyrrolysine amino acid for its direct incorporation into proteins inside living cells. In: Chemistry-A European Journal, Vol. 21, No. 21: pp. 7701-7704


We here report the construction of an E. coli expression system able to manufacture an unnatural amino acid by an artificial biosynthesis. This can be orchestrated with incorporation into protein by amber stop codon suppression inside a living cell. In our case an alkyne bearing pyrrolysine amino acid was biosynthesized and incorporated site-specifically allowing orthogonal double protein labeling.