Acinetobacter o-oligosaccharyltransferases and uses thereof
The present application provides methods and uses of O-oligosaccharyltransferase (O-OTases) for generating vaccines. In particular, the present application provides a method of synthesizing a glycoprotein comprising glycosylation of pilin-like protein ComP using a PglL ComP O-OTase. Uses of glycoproteins synthesized by glycosylating ComP using PglL ComP O-OTase, particularly for the preparation of vaccines and the like, including a vaccine to Streptococcus , is also provided.
1. A method of glycosylating a protein in a bacterial cell, the method comprising glycosylating a fusion protein comprising ComP, with an N-glycan in a bacterial cell, using a PglL ComP O-oligosaccharyltransferase.
2. The method of claim 1 comprising expressing PglL ComP O-oligosaccharyltransferase, the ComP fusion protein, and the N-glycan together in the bacterial cell.
3. The method of claim 1 , wherein the ComP fusion protein comprises an adjuvant or carrier.
4. The method of claim 1 , wherein the bacterial cell is Acinetobacter or E. coli.
5. The method of claim 1 , wherein the N-glycan is derived from Campylobacter.
6. The method of claim 5 , wherein the N-glycan is a Campylobacter jejuni N-heptasaccharide.
7. The method of claim 4 , wherein the Acinetobacter is A. baylyi, A. baumannii, A. nosocomialis , or A. calcoaceticus.
8. A conjugate vaccine comprising the glycosylated ComP fusion protein produced in accordance with the method of claim 1 .
9. The method of claim 4 , wherein the bacterial cell is E. coli.
10. The conjugate vaccine of claim 8 , wherein the vaccine comprises an adjuvant or carrier.
11. A conjugate vaccine comprising the glycosylated ComP fusion protein produced in accordance with the method of claim 2 .
12. The conjugate vaccine of claim 11 , wherein the vaccine comprises an adjuvant or carrier.
13. A conjugate vaccine comprising the glycosylated ComP fusion protein produced in accordance with the method of claim 5 .
14. The conjugate vaccine of claim 13 , wherein the vaccine comprises an adjuvant or carrier.
15. A conjugate vaccine comprising the glycosylated ComP fusion protein produced in accordance with the method of claim 6 .
16. The conjugate vaccine of claim 15 , wherein the vaccine comprises an adjuvant or carrier.