The global spread of multi- and pan-resistant bacteria has triggered research to identify
novel strategies to fight these pathogens, such as antimicrobial peptides and, more recently, bacteriophages.
In a proof-of-concept study, we have genetically modified lytic T7Select phages targeting
Escherichia coli Rosetta by integrating DNA sequences derived from the proline-rich antimicrobial
peptide, apidaecin. This allowed testing of our hypothesis that apidaecins and bacteriophages can
synergistically act on phage-sensitive and phage-resistant E. coli cells and overcome the excessive cost
of peptide drugs by using infected cells to express apidaecins before cell lysis. Indeed, the addition
of the highly active synthetic apidaecin analogs, Api802 and Api806, to T7Select phage-infected
E. coli Rosetta cultures prevented or delayed the growth of potentially phage-resistant E. coli Rosetta
strains. However, high concentrations of Api802 also reduced the T7Select phage fitness. Additionally,
plasmids encoding Api802, Api806, and Api810 sequences transformed into E. coli Rosetta allowed
the production of satisfactory peptide quantities. When these sequences were integrated into the
T7Select phage genome carrying an N-terminal green fluorescent protein (GFP-) tag to monitor the
expression in infected E. coli Rosetta cells, the GFP–apidaecin analogs were produced in reasonable
quantities. However, when Api802, Api806 and Api810 sequences were integrated into the T7Select
phage genome, expression was below detection limits and an effect on the growth of potentially
phage-resistant E. coli Rosetta strains was not observed for Api802 and Api806. In conclusion, we
were able to show that apidaecins can be integrated into the T7Select phage genome to induce their
expression in host cells, but further research is required to optimize the engineered T7Select phages
for higher expression levels of apidaecins to achieve the expected synergistic effects that were visible
when the T7Select phages and synthetic Api802 and Api806 were added to E. coli Rosetta cultures.
| Identifer | oai:union.ndltd.org:DRESDEN/oai:qucosa:de:qucosa:89051 |
| Date | 16 January 2024 |
| Creators | Ludwig, Tobias, Hoffmann, Ralf, Krizsan, Andor |
| Publisher | MDPI |
| Source Sets | Hochschulschriftenserver (HSSS) der SLUB Dresden |
| Language | English |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, doc-type:article, info:eu-repo/semantics/article, doc-type:Text |
| Rights | info:eu-repo/semantics/openAccess |
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