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Streptomyces albus: A New Cell Factory for Non-Canonical Amino Acids Incorporation into Ribosomally Synthesized Natural Products
journal contribution
posted on 2017-07-31, 00:00 authored by Mariia Lopatniuk, Maksym Myronovskyi, Andriy LuzhetskyyThe incorporation
of noncanonical amino acids (ncAAs) with different
side chains into a peptide is a promising technique for changing the
functional properties of that peptide. Of particular interest is the
incorporation of ncAAs into peptide-derived natural products to optimize
their biophysical properties for medical and industrial applications.
Here, we present the first instance of ncAA incorporation into the
natural product cinnamycin in streptomycetes using the orthogonal
pyrrolysyl-tRNA synthetase/tRNAPyl pair from Methanosarcina
barkeri. This approach allows site-specific incorporation
of ncAAs via the read-through of a stop codon by
the suppressor tRNAPyl, which can carry different pyrrolysine
analogues. Five new deoxycinnamycin derivatives were obtained with
three distinct pyrrolysine analogues incorporated into diverse positions
of the antibiotic. The combination of partial hydrolysis and MS/MS
fragmentation analysis was used to verify the exact position of the
incorporation events. The introduction of ncAAs into different positions
of the peptide had opposite effects on the peptide’s biological
activity.