Catalytic domains of beta(1,4)-galactosyl transferase i having altered donor and acceptor specificities, domains that promote in vitro protein folding, and methods for their use

Abstract

Disclosed are methods and compositions that can be used to synthesize oligosaccharides; mutants of galactosyltransferases having altered donor and acceptor specificity; methods for increasing the immunogenicity of an antigen; and polypeptide stem regions that can be used to promote in vitro folding of polypeptides, such as the catalytic domain from a galactosyltransferase.

1 .- 11 . (canceled) 
     
     
         12 . An isolated catalytic domain from a β(1,4)-galactosyltransferase I that can catalyze formation of a galactose-β(1,4)-N-acetylglucosamine-6-SO 3  bond. 
     
     
         13 . The catalytic domain of  claim 12 , wherein
 (a) the catalytic domain has conservative amino acid exchanges at amino acid positions 279 and 280, or   (b) serine is exchanged for lysine at amino acid position 279, and threonine is exchanged for phenylalanine at amino acid position 280.   
     
     
         14 .- 51 . (canceled) 
     
     
         52 . A kit comprising packaging material, and a β(1,4)-galactosyltransferase I catalytic domain that: can catalyze formation of a galactose-β(1,4)-N-acetylglucosamine-6-SO 3  bond. 
     
     
         53 . The kit of  claim 52 , further comprising a donor. 
     
     
         54 . The kit of  claim 53 , wherein the donor is selected from the group consisting of UDP-galactose, UDP-mannose, UDP-N-acetylglucosamine, UDP-glucose, GDP-mannose, UDP-N-acetylgalactosamine, UDP-glucuronic acid, GDP-Fucose, and CMP-N-acetylneuraminic acid.