Enzymatic method for preparation of UDP-GlcNAc
AssigneeMax-Planck-Gesellschaft zur Forderung der Wissenschaften e.V.
InventorsReza Mahour, Thomas F. T. Rexer
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Abstract
The present invention relates to an enzyme-catalyzed process for producing UDP-N-acetyl-α-D-glucosamine (UDP-GlcNAc) from low-cost substrates uridine monophosphate and N-acetyl-D glucosamine in a single reaction mixture with immobilized or preferably co-immobilized enzymes. Uridine may be used as starting material instead of uridine monophosphate as well. Further, the process may be adapted to produce GlcNAcylated molecules and biomolecules including saccharides, particularly human milk oligosaccharides (HMO), proteins, peptides, glycoproteins, particularly antibodies, or glycopeptides, and bioconjugates, particularly carbohydrate conjugate vaccines and antibody-drug conjugates.
Claims16
A method for producing uridine 5′-diphospho-N-acetyl-α-D-glucosamine comprising:
- providing a solution comprising
- (i) uridine monophosphate and N-acetyl-D-glucosamine represented by the following formulae
- (ii) polyphosphate, and adenosine triphosphate;
- and
- providing a set of enzymes comprising a glucose-1-phosphate uridylyltransferase, an N-acetylhexosamine kinase, a polyphosphate kinase, and a uridine monophosphate kinase;
- A) producing uridine 5′-diphospho-N-acetyl-α-D-glucosamine from uridine monophosphate and N-acetylglucosamine in the presence of the set of enzymes, polyphosphate, and adenosine triphosphate,
- wherein the set of enzymes is covalently or adsorptively immobilized on a reusable, mechanically stable solid support.
Dependencies are inferred from the claim text.