Multimeric antimicrobial peptide complex which is displayed on cell surface

Abstract

The present invention provides an antimicrobial peptide polymer comprising at least one monomer which is digested by pepsin, a multimeric antimicrobial peptide complex comprising the polymer and a cell surface anchoring motif linked to the polymer, an antimicrobial microorganism displaying the multimeric antimicrobial peptide complex, an antimicrobial composition comprising the same, a method of treating an infectious disease caused by bacteria, yeast or fungi by administering the antimicrobial composition, and a method for producing the antimicrobial microorganism. According to the invention, living microorganisms displaying an antimicrobial peptide on the cell surface thereof may be administered in vivo without having to lyse the microbial cell and isolate and purify the antimicrobial peptide, so that the antimicrobial peptide exhibits antimicrobial activity. Thus, the antimicrobial peptide may be produced at significantly reduced costs so that it may have widespread use.

1 . An isolated multimeric antimicrobial peptide complex comprising an antimicrobial peptide polymer and a cell surface anchoring motif linked to the polymer, wherein the antimicrobial peptide polymer comprises at least one monomer represented by the following formula 1 or 2:
   N-terminus-[antimicrobial peptide-amino acid linker digested by pepsin]-C-terminus,  Formula 1
     N-terminus-[amino acid linker digested by pepsin-antimicrobial peptide]-C-terminus,  Formula 2
   wherein the antimicrobial peptide does not comprise the amino acid used in the linker.   
     
     
         2 . The isolated multimeric antimicrobial peptide complex of  claim 1 , wherein the antimicrobial peptide has any one of amino acid sequences represented by SEQ ID NOS: 9 to 24. 
     
     
         3 . The isolated multimeric antimicrobial peptide complex of  claim 1 , wherein the amino acid linker digested by pepsin is selected from the group consisting of leucine, phenylalanine and tyrosine. 
     
     
         4 . The isolated multimeric antimicrobial peptide complex of  claim 1 , wherein the cell surface anchoring motif is linked to the N-terminus of the polymer. 
     
     
         5 . The isolated multimeric antimicrobial peptide complex of  claim 1 , wherein the cell surface anchoring motif is an outer membrane protein. 
     
     
         6 . The isolated multimeric antimicrobial peptide complex of  claim 5 , wherein the outer membrane protein is selected from the group consisting of an  E. coli  outer membrane protein OmpA, an  E. coli  outer membrane protein OmpA linked to the leader sequence of  E. coli  lipoprotein, an  E. coli  outer membrane protein OmpS, an  E. coli  outer membrane protein LamB, an  E. coli  outer membrane protein PhoE, an  E. coli  outer membrane protein OmpC, an  E. coli  outer membrane protein Fad L, a  Salmonella  outer membrane protein OmpC, and a  Pseudomonas  outer membrane protein OprF. 
     
     
         7 . The isolated multimeric antimicrobial peptide complex of  claim 6 , wherein the  E. coli  outer membrane protein OmpA linked to the leader sequence of  E. coli  lipoprotein has an amino acid sequence represented by SEQ ID NO: 8. 
     
     
         8 . The isolated multimeric antimicrobial peptide complex of  claim 1 , wherein the antimicrobial peptide has an amino acid sequence represented by SEQ ID NO: 9, the amino acid linker digested by pepsin is leucine, and the cell surface anchoring motif has an amino acid sequence represented by SEQ ID NO: 8. 
     
     
         9 . An isolated antimicrobial peptide polymer comprising at least one monomer represented by the following formula 1 or 2:
   N-terminus-[antimicrobial peptide-amino acid linker digested by pepsin]-C-terminus,  Formula 1
     N-terminus-[amino acid linker digested by pepsin-antimicrobial peptide]-C-terminus,  Formula 2
   wherein the antimicrobial peptide does not comprise the amino acid used in the linker.   
     
     
         10 . A polynucleotide encoding the isolated multimeric antimicrobial peptide complex of  claim 1 . 
     
     
         11 . A polynucleotide encoding the isolated antimicrobial peptide polymer of  claim 9 . 
     
     
         12 . The polynucleotide of  claim 10 , wherein the polynucleotide has any one of nucleotide sequences represented by SEQ ID NOS: 25 to 27. 
     
     
         13 . The polynucleotide of  claim 11 , wherein the polynucleotide has any one of nucleotide sequences of SEQ ID NOS: 28 to 30. 
     
     
         14 . A recombinant vector comprising the polynucleotide of  claim 10 . 
     
     
         15 . An antimicrobial microorganism transformed with the recombinant vector of  claim 14  to display a multimeric antimicrobial peptide complex on the cell surface thereof. 
     
     
         16 . (canceled) 
     
     
         17 . An antimicrobial composition comprising the isolated multimeric antimicrobial peptide complex of  claim 1  as an active ingredient. 
     
     
         18 . An antimicrobial composition comprising the isolated antimicrobial peptide polymer of  claim 9  as an active ingredient. 
     
     
         19 . (canceled) 
     
     
         20 . The antimicrobial composition of  claim 17 , wherein the composition is a pharmaceutical composition or an over-the-counter (OTC) drug composition. 
     
     
         21 . A method for producing an antimicrobial microorganism displaying a multimeric antimicrobial peptide complex on the cell surface thereof, comprising (a) preparing a recombinant vector comprising a polynucleotide encoding the isolated multimeric antimicrobial peptide complex of  claim 1 ; (b) introducing the recombinant vector into a host cell to obtain a transformant; and (c) culturing the transformant to induce the expression of the multimeric antimicrobial peptide complex. 
     
     
         22 . A method for treating an infectious disease caused by pathogenic bacteria, yeast or fungi, comprising administering the antimicrobial composition of  claim 17  to a subject having the infectious disease. 
     
     
         23 . (canceled)