FlmG-dependent soluble protein O-glycosylation systems in bacteria

A polynucleotide flmG which encodes a Flagellin Modification Protein (FlmG) having glycosyltransferase activity and being selected from the group consisting of a), b), and c):

• a) the polynucleotide composed of SEQ ID NO:
• 26 or the polynucleotide encoding the protein composed of SEQ ID NO:
• 27;
• b) a polynucleotide having 95% or more sequence identity with the polynucleotide sequence of SEQ ID NO:
• 26 or with the polynucleotide sequence encoding the protein composed of SEQ ID NO:
• 27, and which encodes a protein having activity that transfers a soluble monosaccharide to the hydroxyl group on a threonine residue of a soluble acceptor protein of interest;
• and
• c) a polynucleotide that encodes a protein that has an amino acid sequence having 95% or more sequence identity with the amino acid sequence of SEQ ID NO:
• 27 and has activity that transfers a soluble monosaccharide to the hydroxyl group on a threonine residue of the soluble acceptor protein of interest;
• wherein glycosylation is an O-glycosylation of the soluble acceptor protein of interest in the presence of the monosaccharide which is performed within the cytoplasm of a bacterial Gram-negative cell;
• and
• wherein the soluble acceptor protein of interest is selected from the group consisting of:
• i. a flagellin;
• ii. a protein selected from the group consisting of Alpha-1-Antitrypsin, Interferon-beta, insulin, and an antimicrobial peptide;
• and
• iii. a flagellin fused to another soluble protein of interest.