Endolysin OBPgpLYS
The present invention relates to a polypeptide with an amino acid sequence according to SEQ ID NO: 1 and fragments or derivatives thereof. The present invention further relates to fusion proteins comprising said polypeptide and an additional peptide stretch fused to said polypeptide at the N- or C-terminus. Moreover, the present invention relates to nucleic acid molecules encoding said polypeptide or fusion protein, vectors comprising said nucleic acid molecules and host cells comprising either said nucleic acid molecules or said vectors. In addition, the present invention relates to said polypeptide or fusion protein for use as a medicament, in particular for the treatment or prevention of Gram-negative bacterial infections, as diagnostic means, as cosmetic substance or as sanitizing agent. The present invention also relates to the use of said polypeptide or fusion protein for 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. Furthermore, the present invention relates to a pharmaceutical composition comprising said polypeptide or fusion protein.
1. A fusion polypeptide comprising (i) the amino acid sequence of SEQ ID NO: 1, or a fragment thereof, wherein the fragment comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 4 and 5, or comprises the amino acid sequences of SEQ ID NO: 4 and 5, and (ii) a heterologous polypeptide.
2. The fusion polypeptide according to claim 1 , further comprising a tag.
3. The fusion polypeptide according to claim 2 , wherein said fusion polypeptide comprises the amino acid sequence of SEQ ID NO: 47.
4. A fusion protein comprising (i) a polypeptide comprising the amino acid sequence of SEQ ID NO: 1, or a fragment thereof, wherein the fragment comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 4 and 5, or comprises the amino acid sequences of SEQ ID NO: 4 and 5, and (ii) a heterologous peptide fused to said polypeptide at the N- or C-terminus, wherein said heterologous peptide is a cationic peptide, polycationic peptide, amphipathic peptide, sushi peptide, defensin, hydrophobic peptide and/or an antimicrobial peptide.
5. The fusion protein according to claim 4 , wherein said heterologous peptide comprises 5 to 100 amino acid residues.
6. The fusion protein according to claim 4 , wherein said cationic and/or polycationic peptide comprises at least one amino acid residue selected from the group consisting of arginine, histidine and lysine residues.
7. The fusion protein according to claim 4 , wherein the amphipathic peptide comprises at least one positively charged amino acid residue selected from the group consisting of lysine, arginine and histidine residues, combined to at least one hydrophobic amino acid residue selected from the group consisting of valine, isoleucine, leucine, methionine, phenylalanine, tryptophan, cysteine, alanine, tyrosine, histidine, threonine, serine, proline and glycine residues.
8. The fusion protein according to claim 4 , wherein the heterologous peptide comprises an amino acid sequence selected from the group consisting SEQ ID NO: 19-39 and KRK.
9. The fusion protein according to claim 4 , wherein said fusion protein comprises an amino acid sequence selected from the group consisting SEQ ID NO: 43 and 49.
10. A pharmaceutical or cosmetic composition comprising the fusion polypeptide according to claim 1 .
11. A pharmaceutical or cosmetic composition comprising the fusion protein of claim 4 .
12. The fusion polypeptide according to claim 2 , wherein the tag comprises a His 6 -tag.
13. The fusion protein according to claim 5 , wherein said heterologous peptide comprises 5 to 50 amino acid residues.
14. The fusion protein according to claim 13 , wherein said heterologous peptide comprises 5 to 30 amino acid residues.
15. The fusion protein according to claim 6 , wherein at least 70% of the amino acid residues comprised in said heterologous peptide are arginine, histidine and/or lysine residues.
16. The fusion protein according to claim 15 , wherein at least 70% of the amino acid residues comprised in said heterologous peptide are arginine and/or lysine residues.
17. The fusion protein according to claim 7 , wherein at least 70% of the amino acid residues in said amphipathic peptide are either arginine or lysine residues and at least 30% of the amino acid residues in said amphipathic peptide are valine, isoleucine, leucine, methionine, phenylalanine, tryptophan, cysteine, alanine, tyrosine, histidine, threonine, serine, proline or glycine residues.
18. The fusion protein according to claim 4 , wherein the heterologous peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 6, 8, 10, 11, and 15.
19. The fusion protein according to claim 4 , wherein the heterologous peptide comprises the amino acid sequence of SEQ ID NO: 14.
20. The fusion protein according to claim 4 , wherein the heterologous peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 7, 9, 12, 16, and 17.
21. The fusion protein according to claim 4 , wherein the heterologous peptide comprises the amino acid sequence of SEQ ID NO: 13.
22. The fusion protein according to claim 4 , wherein the heterologous peptide comprises the amino acid sequence of SEQ ID NO: 18.
23. A fusion polypeptide comprising (i) the amino acid sequence of SEQ ID NO: 86 or 87 and (ii) a heterologous polypeptide.
24. A fusion protein comprising (i) a polypeptide comprising the amino acid sequence of SEQ ID NO: 86 or 87, and (ii) a heterologous peptide fused to said polypeptide at the N- or C-terminus, wherein said heterologous peptide is a cationic peptide, polycationic peptide, amphipathic peptide, sushi peptide, defensin, hydrophobic peptide and/or an antimicrobial peptide.