| abstract |
One preferred embodiment of the present invention provides a GalK variant comprising a R28, E34, D37, D174, Y233, C339, Y371, Y371H, M173, M173L, Y371H-M173L and C353 mutation. The GalK variants display catalytic activity toward a variety of D or L sugars. Another preferred embodiment provides method of phosphorylating sugars comprising the step of incubating a nucleotide triphosphate (NTP) and a D or L sugar in the presence of a GalK variant such that a sugar phosphate is produced. This sugar phosphate may be further incubated with a nucleotidylyltransferase, such that a NDP-sugar is produced. The NDP-sugar may be further incubated with a biomolecule to produce a glycosylated biomolecule. |