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 Pg1L ComP O-OTase. Uses of glycoproteins synthesized by glycosylating ComP using Pg1L ComP O-OTase, particularly for the preparation of vaccines and the like, including a vaccine to Streptococcus , is also provided.
1. A bacterial cell expressing a PglLcomP O-oligosacharyltransferase, a fusion protein comprising ComP, and a polysaccharide containing glucose at its reducing end.
2. The bacterial cell of claim 1 wherein the cell also comprises the ComP fusion protein glycosylated with the polysaccharide.
3. The bacterial cell of claim 1 , wherein the bacterial cell is Acinetobacter or E. coli.
4. The bacterial cell of claim 3 , wherein the Acinetobacter is A. baylyi, A. baumannii, A. nosocomialis , or A. calcoaceticus.
5. The bacterial cell of claim 3 , wherein the bacterial cell is E. coli.
6. The bacterial cell of claim 1 , wherein the polysaccharide is a bacterial capsular polysaccharide.
7. The bacterial cell of claim 6 , wherein the capsular polysaccharide is a Streptococcus capsule.
8. The bacterial cell of claim 6 , wherein the capsular polysaccharide is a Streptococcus pneumoniae capsule.
9. The bacterial cell of claim 2 , wherein the bacterial cell is Acinetobacter or E. coli.
10. The bacterial cell of claim 9 , wherein the Acinetobacter is A. baylyi, A. baumannii, A. nosocomialis , or A. calcoaceticus.
11. The bacterial cell of claim 9 , wherein the bacterial cell is E. coli.
12. The bacterial cell of claim 2 , wherein the polysaccharide is a bacterial capsular polysaccharide.
13. The bacterial cell of claim 12 , wherein the capsular polysaccharide is a Streptococcus capsule.
14. The bacterial cell of claim 12 , wherein the capsular polysaccharide is a Streptococcus pneumoniae capsule.
15. A bacterial cell comprising a glycosylated fusion protein comprising ComP, wherein the ComP fusion protein is glycosylated with a polysaccharide containing glucose at its reducing end.
16. The bacterial cell of claim 15 , wherein the bacterial cell is Acinetobacter or E. coli.
17. The bacterial cell of claim 16 , wherein the bacterial cell is E. coli.
18. The bacterial cell of claim 15 , wherein the polysaccharide is a bacterial capsular polysaccharide.
19. The bacterial cell of claim 18 , wherein the capsular polysaccharide is a Streptococcus capsule.
20. The bacterial cell of claim 18 , wherein the capsular polysaccharide is a Streptococcus pneumoniae capsule.