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1Department of Biocatalysis, Institute of Chemistry,Technische Universität Berlin, 2Department of Bioenergetics, Institute of Chemistry,Technische Universität Berlin, 3Molecular Genetics Group, Groningen Biomolecular Sciences and Biotechnology Institute, Department of Molecular Genetics,University of Groningen
Chapters
- 00:05Title
- 01:16Expression of Recombinant Nisin with Incorporation of Proline Analogs by SPI
- 04:19Antimicrobial Activity Test
- 07:37Fluorescence Microscopy
- 09:02Results: Fluorescence Microscopy of Recombinant Nisin Effects on Gram-positive Cells
- 10:38Conclusion
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.
