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 a bacterial capsular polysaccharide containing glucose at its reducing end, using a PglL comP O-oligosaccharyltransferase.
2. The method of claim 1 comprising expressing PglL comP O-oligosccharyltransferase, the ComP fusion protein, and the capsular polysaccharide 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 capsular polysaccharide is a Streptococcus capsule.
6. The method of claim 1 , wherein the capsular polysaccharide
is a Streptococcus pneumoniae capsule.
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 conjugate vaccine of claim 8 , wherein the capsular polysaccharide is a Streptococcus capsule and the vaccine is a Streptococcus vaccine.
10. A glycosylated fusion protein comprising ComP, wherein the fusion protein is glycosylated with a bacterial capsular polysaccharide containing glucose at its reducing end.
11. The method of claim 4 , wherein the bacterial cell is E. coli.
12. The conjugate vaccine of claim 8 , wherein the capsular polysaccharide is a Streptococcus pneumoniae capsule and the vaccine is a Streptococcus pneumoniae vaccine.
13. The conjugate vaccine of claim 8 , wherein the vaccine comprises an adjuvant or carrier.
14. The glycosylated fusion protein of claim 10 , wherein the capsular polysaccharide is from Streptococcus.
15. The glycosylated fusion protein of claim 10 , wherein the capsular polysaccharide is from Streptococcus pneumoniae.
16. A conjugate vaccine comprising the glycosylated ComP fusion protein of claim 10 .
17. The conjugate vaccine of claim 16 , wherein the capsular polysaccharide is a Streptococcus capsule and the vaccine is a Streptococcus vaccine.
18. The conjugate vaccine of claim 16 , wherein the capsular polysaccharide is a Streptococcus pneumoniae capsule and the vaccine is a Streptococcus pneumoniae vaccine.
19. The conjugate vaccine of claim 16 , wherein the vaccine comprises an adjuvant or carrier.