Surface expression method of peptides P5 and Anal3 using the gene encoding poly-gamma-glutamate synthetase
The present invention relates to a method for expressing each of peptide antibiotics P5 3 and Ana13 35 having amphiphilicity and showing antibacterial, antifungal and anticancer activities 61, 63, 65, 67, 69, 71, on the microbial surface, using a vector containing outer membrane protein genes (pgsBCA) that are derived from Bacillus sp. strains and involved in the synthesis of poly-gamma-glutamate. Moreover, the present invention relates to lactic acid-forming bacteria having each of the peptide antibiotics P5 15 and Ana13 43 expressed on their surface, and the use thereof. According to the present invention, the peptide antibiotics can be expressed on the surface of various microorganisms transformed with the surface expression vectors. The inventive method for the surface expression of the peptide antibiotics allows the peptide antibiotics to be mass-produced without a purification process. Thus, the inventive method has very high industrial applicability. Further, the present invention can be applied to other peptide antibiotics besides P5 3 and Ana13 35.
1. A vector for the surface expression of antibiotics, which comprises:
one or more than two genes selected from the group consisting of pgsB, pgsC and pgsA, said genes encoding a poly-gamma-glutamate synthetase complex; and
a gene encoding P5 peptide having antibacterial, antifungal and anticancer activities fused with said gene encoding a poly-gamma-glutamate synthetase complex, wherein P5 peptide is encoded by the base sequence of SEQ ID NO: 4.
2. The vector according to claim 1 , wherein said pgsB, pgsC and pgsA genes have the base sequences described in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, respectively.
3. The vector according to claim 1 , wherein the vector contains the pgsA gene among the genes encoding the poly-gamma-glutamate synthetase complex.
4. The vector according to claim 1 , said vector is pHCE1LB:pgsA-P5 for the surface expression of antibiotics, which expresses said antibiotic on the surface of gram-negative and gram positive bacteria.
5. A microorganism transformed with the vector of claim 1 .
6. E. Coli (KCTC 10350BP) transformed with the vector pHCE1LB:pgsA-P5 of claim 4 .
7. A lactic acid-forming bacteria transformed with the vector of claim 1 .
8. A pharmaceutical composition and suspension of the same for antibacterial, antifungal or anticancer application, which comprises, as an active ingredient, the lactic acid-forming bacteria according to claim 7 and having the peptide antibiotic P5 expressed on their surface.
9. The pharmaceutical composition according to claim 8 , wherein said active ingredient is heat-treated.