Peptides for inducing bacteriocin synthesis and methods to identify and/or select and/or optimize the same
Inventors
Mignolet, Johann • Hols, Pascal • LEDESMA GARCÍA, Laura
Assignees
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Publication Number
US-12234299-B2
Publication Date
2025-02-25
Expiration Date
Abstract
Described herein is a peptide or peptidomimetic with a length of at least 6 residues comprising, consisting essentially of, or consisting of the sequence motif Xaa1-Trp-Xaa2-Xaa3-Xaa4-Xaa5 (SEQ ID NO:1), wherein: Xaa1 represents an aromatic residue (Phe, Tyr, Trp, His), Cys or Ser; Xaa2, Xaa3 and Xaa4 represent any residue; and Xaa5 represents Gly, lie or Val.
Core Innovation
The invention relates to a peptide of at least 6 residues that induces bacteriocin production in a microbial organism. The peptide comprises a sequence motif selected from Phe-Trp-Leu-Val-Leu-Gly (SEQ ID NO: 2), Ala-Phe-Trp-Leu-Ile-Leu-Gly (SEQ ID NO: 3), and Pro-Phe-Trp-Leu-Ile-Leu-Gly (SEQ ID NO: 7). The motif is located at the C-terminus of the peptide or is located so that it is followed by at most 1, 2, 3, 4 or 5 additional C-terminal residues.
The invention further covers a pro-polypeptide comprising a peptide sequence selected from the specified motif sequences, where one or more cleavage sites separate the peptide sequence from the rest of the pro-polypeptide. Release of the active peptide from the pro-polypeptide is described as occurring by peptide hydrolysis.
The invention also relates to a regulatory system in Streptococcus salivarius HSISS4 in which Rgg-family paralogs ComR, ScuR, and SarF control competence and bacteriocin production. In this system, ScuR regulates sptA/sptB and bacteriocin promoters while maintaining competence off, and competence and bacteriocin functions are decoupled.
Documented results indicate that promoter-controlled reporter outputs show synthetic peptides activate ScuR/SarF-dependent light/bacteriocin output without inducing ComX/competence genes. A conserved tryptophan in the peptide motif is required for activation of ScuR and/or SarF, and engineered promoter nucleotide topology can switch pheromone selectivity.
Claims Coverage
The independent claims are centered on motif-containing peptides and pro-polypeptides, with coverage for C-terminal motif placement, limited additional C-terminal residues, and cleavage sites that separate the motif-bearing sequence from the remainder of the pro-polypeptide. In total, the claim set defines these motif, placement, and cleavage features as the core inventive concepts.
C-terminal Trp-centered bacteriocin-inducing peptide motif
A peptide able to induce bacteriocin production in a microbial organism with a length of at least 6 residues, comprising a sequence motif selected from Phe-Trp-Leu-Val-Leu-Gly (SEQ ID NO: 2), Ala-Phe-Trp-Leu-Ile-Leu-Gly (SEQ ID NO: 3), and Pro-Phe-Trp-Leu-Ile-Leu-Gly (SEQ ID NO: 7), wherein the sequence motif is located at the C-terminus of said peptide or is located so that it is followed by at most 1, 2, 3, 4 or 5 additional C-terminal residues.
Pro-polypeptide with cleavage sites releasing C-terminal motif peptide
A pro-polypeptide comprising a peptide sequence comprising an amino acid sequence selected from Phe-Trp-Leu-Val-Leu-Gly (SEQ ID NO: 2), Ala-Phe-Trp-Leu-Ile-Leu-Gly (SEQ ID NO: 3), and Pro-Phe-Trp-Leu-Ile-Leu-Gly (SEQ ID NO: 7), wherein the amino acid sequence is located at the C-terminus of said peptide or is located so that it is followed by at most 1, 2, 3, 4 or 5 additional C-terminal residues; and one or more cleavage sites, wherein the one or more cleavage sites separates the peptide sequence from the rest of the pro-polypeptide.
The independent claims collectively require a peptide or pro-polypeptide with a specified Trp-centered motif chosen from SEQ ID NO: 2, 3, or 7, motif placement at the C-terminus or within a limit of additional C-terminal residues, and, in the pro-polypeptide claim, one or more cleavage sites that separate the motif-bearing peptide from the remainder of the pro-polypeptide.
Stated Advantages
Induces bacteriocin production in a microbial organism.
Synthetic peptides activate ScuR/SarF-dependent light/bacteriocin output without inducing ComX/competence genes.
ScuR maintains competence off while regulating sptA/sptB and bacteriocin promoters.
Documented Applications
Using synthetic peptides to activate ScuR/SarF-dependent light/bacteriocin output without inducing ComX/competence genes in Streptococcus salivarius HSISS4.
A regulatory-system context in Streptococcus salivarius HSISS4 where ComR, ScuR, and SarF control competence and bacteriocin production.
Induction of bacteriocin production using the motif-containing peptide, including formats such as culture medium and solvent-containing compositions.
Use of a genetically modified microbial organism that produces, or produces and secretes, the motif-containing peptide.
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