Patent
The present invention is directed to sialytransferases, such as SiaA sialytransferases isolated from Haemophilus influenzae . Further provided herein are methods for producing sialylated lipooligosaccharides, vaccines, and host cells and systems for the production of sialylated lipooligosaccharides.
1. An isolated polynucleotide comprising SEQ ID NO:1.
2. The isolated polynucleotide comprising nucleotides 2538-3452 of SEQ ID NO:1.
3. An expression cassette comprising the isolated polynucleotide of claim 1 or claim 2 .
4. An isolated cell comprising the expression cassette of claim 3 .
5. The cell of claim 4 , further comprising an expression cassette comprising a nucleotide sequence encoding SEQ ID NO:4.
6. An isolated lipooligosaccharide (LOS) production cell comprising:
(i) a nucleic acid encoding SEQ ID NO:4;
(ii) the expression cassette of claim 3 ;
(iii) an acceptor molecule; and
(iv) a substrate for a sialyltransferase.
7. The production cell of claim 6 , wherein the cell is a gram negative bacterium.
8. The production cell of claim 7 , wherein the bacterium is Haemophilus influenzae or Escherichia coli.
9. An isolated polynucleotide comprising a nucleic acid sequence, wherein the nucleic acid sequence encodes SEQ ID NO:3.
10. An expression cassette comprising the isolated polynucleotide of claim 9 .
11. An isolated cell comprising the expression cassette of claim 10 .
12. The cell of claim 11 , further comprising an expression cassette comprising a nucleotide sequence encoding SEQ ID NO:4.
13. An isolated lipooligosaccharide (LOS) production cell comprising:
(i) a nucleic acid encoding SEQ ID NO:4;
(ii) the expression cassette of claim 10 ;
(iii) an acceptor molecule; and
(iv) a substrate for a sialyltransferase.
14. The production cell of claim 13 , wherein the cell is a gram negative bacterium.
15. The production cell of claim 14 , wherein the bacterium is Haemophilus influenzae or Escherichia coli.