The protocol presents the Escherichia coli-based selective pressure incorporation of non-canonical amino acids (ncAAs) into the lactococcal antimicrobial peptide nisin. Its properties can be changed during recombinant expression via substitution with desired ncAAs in defined growth media. Resulting changes in bioactivity are mapped by growth inhibition assays and fluorescence microscopy.
Nickling, J. H., Baumann, T., Schmitt, F., Bartholomae, M., Kuipers, O. P., Friedrich, T., Budisa, N. Antimicrobial Peptides Produced by Selective Pressure Incorporation of Non-canonical Amino Acids. J. Vis. Exp. (135), e57551, doi:10.3791/57551 (2018).