Antimicrobial agents
The application relates to antimicrobial agents against Gram-negative bacteria, in particular to fusion proteins composed of an enzyme having the activity of degrading the cell wall of Gram-negative bacteria and a peptide stretch fused to the enzyme at the N- or C-terminus, as well as pharmaceutical compositions thereof. Moreover, it relates to nucleic acid molecules encoding such a fusion protein, vectors carrying the nucleic acid molecules and host cells transformed with either the nucleic acid molecules or the vectors. In addition, it relates to such a fusion protein for use as a medicament, in particular for the treatment or prevention of Gram-negative bacterial infections, as diagnostic means or as cosmetic substance. The application also relates to the treatment or prevention of Gram-negative bacterial contamination of foodstuff, of food processing equipment, of food processing plants, of surfaces coming into contact with foodstuff, of medical devices, of surfaces in hospitals and surgeries.
1. A fusion protein comprising an endolysin having Gram-negative bacteria cell wall degrading activity and an amphipathic peptide segment fused to the endolysin at the N- or C-terminus or at both termini, wherein the amphipathic peptide segment comprises SMAP-29 having the sequence of SEQ ID NO: 11.
2. The fusion protein according to claim 1 , wherein said fusion protein is formulated in a pharmaceutical carrier.
3. The fusion protein according to claim 1 , wherein said endolysin is an endolysin of the Pseudomonas aeruginosa phages ΦKZ or EL.
4. The fusion protein according to claim 1 , wherein said endolysin is an endolysin having an amino acid sequence according to SEQ ID NO: 25, 22, 24, 2, 3, 4, 5, 18, 20, 21, or 23.
5. The fusion protein according to claim 1 , wherein said fusion protein further comprises an amino acid residue on the N-terminus that is not naturally found in said endolysin or said amphipathic peptide segment.
6. The fusion protein according to claim 1 , wherein said fusion protein comprises a tag or additional protein on the C- and/or N-terminus.
7. The fusion protein according to claim 6 , wherein said tag or additional protein is linked to the fusion protein by one or more additional amino acid residues.
8. The fusion protein according to claim 1 , wherein said peptide segment is linked to the fusion protein by one or more additional amino acid residues.
9. The fusion protein according to claim 1 , wherein said fusion protein comprises an amino acid sequence according to SEQ ID NO: 46, 82, 95, or 99.
10. An isolated nucleic acid molecule encoding a fusion protein according to claim 1 .
11. The nucleic acid of claim 10 , further comprised within a vector.
12. A host cell comprising a nucleic acid molecule encoding a fusion protein according to claim 1 .
13. A method of treating a Gram-negative bacterial infection comprising contacting a subject or sample having said infection with a fusion protein according to claim 1 .
14. A method for disinfecting a surface or article or preventing contamination comprising contacting said surface or article with a fusion protein according to claim 1 .