Glycosyltransferase engineering for chemoenzymatic total synthesis of gangliosides

Abstract

Described herein are Campylobacter jejuni β1-4GalNAcT (CjCgtA) variants comprising a polypeptide having at least 80% identity to the amino acid sequence set forth in SEQ ID NO: 1 or SEQ ID NO: 2. Also described herein are Campylobacter jejuni β1-3-galactosyltransferase (CjCgtB) variants comprising a polypeptide having at least 80% identity to the amino acid sequence set forth in SEQ ID NO: 3. Optionally, in the CjCgtA and CjCgtB variants, the N-terminus of the polypeptide is fused to a maltose binding protein. Further described herein is a method for preparing a glycosylated molecule, comprising: forming a reaction mixture comprising (i) a glycosylation donor comprising a sugar component; (ii) a glycosylation acceptor comprising a sphingosine moiety; and (iii) a glycosyltransferase, wherein the glycosyltransferase is a CjCgtA variant as described herein or a CjCgtB variant as described herein; and maintaining the reaction mixture under conditions suitable for forming a glycosylated molecule.

Dependencies are inferred from the claim text.