title: Synthesis of trypsin-resistant variants of the Listeria-active bacteriocin salivaricin P.
abstract: Two-component salivaricin P-like bacteriocins have demonstrated potential as antimicrobials capable of controlling infections in the gastrointestinal tract (GIT). The anti-Listeria activity of salivaricin P is optimal when the individual peptides Sln1 and Sln2 are added in succession at a 1:1 ratio. However, as degradation by digestive proteases may compromise the functionality of these peptides within the GIT, we investigated the potential to create salivaricin variants with enhanced resistance to the intestinal protease trypsin. A total of 11 variants of the salivaricin P components, in which conservative modifications at the trypsin-specific cleavage sites were explored in order to protect the peptides from trypsin degradation while maintaining their potent antimicrobial activity, were generated. Analysis of these variants revealed that eight were resistant to trypsin digestion while retaining antimicrobial activity. Combining the complementary trypsin-resistant variants Sln1-5 and Sln2-3 resulted in a MIC(50) of 300 nM against Listeria monocytogenes, a 3.75-fold reduction in activity compared to the level for wild-type salivaricin P. This study demonstrates the potential of engineering bacteriocin variants which are resistant to specific protease action but which retain significant antimicrobial activity.
authors.affiliation: Teagasc, Moorepark Food Research Centre, Fermoy, County Cork, Ireland.
authors.forename: Eileen F
authors.initials: EF
authors.lastname: O'Shea
authors.forename: Paula M
authors.initials: PM
authors.lastname: O'Connor
authors.forename: Paul D
authors.initials: PD
authors.lastname: Cotter
authors.forename: R Paul
authors.initials: RP
authors.lastname: Ross
authors.forename: Colin
authors.initials: C
authors.lastname: Hill
mesh_terms: D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology; D001430:Bacteriocins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D008089:Listeria monocytogenes / Q000187:drug effects*; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D014357:Trypsin / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D001430:Bacteriocins; D011994:Recombinant Proteins; D014357:Trypsin