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Orchestrating the Biosynthesis of an Unnatural Pyrrolysine Amino Acid for Its Direct Incorporation into Proteins Inside Living Cells
Author(s) -
Ehrlich Michael,
Gattner Michael J.,
Viverge Bastien,
Bretzler Johanna,
Eisen David,
Stadlmeier Michael,
Vrabel Milan,
Carell Thomas
Publication year - 2015
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201500971
Subject(s) - biosynthesis , amino acid , biochemistry , protein biosynthesis , chemistry , cell , living cell , biology , enzyme , microbiology and biotechnology
We here report the construction of an E. coli expression system able to manufacture an unnatural amino acid by 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.