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AssigneeMax-Planck-Gesellschaft zur Forderung der Wissenschaften e.V.
InventorsThomas F.T. Rexer, Reza Mahour
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The present invention relates to a method for producing cytidine 5′-monophospho-N-acetyl-neuraminic acid (CMP-Neu5Ac, 1) from low-cost substrates N-acetyl-D-glucosamine (GlcNAc), pyruvate, cytidine and polyphosphate in a single reaction mixture with a set of optionally immobilized or optionally co-immobilized enzymes comprising N-acylglucoamine 2-epimerase (AGE), an N-acetylneuraminate lyase (NAL), an N-acylneuraminate cytidylyltransferase (CSS), a uridine kinase (UDK), a uridine monophosphate kinase and a polyphosphate kinase 3 (PPK3). Further, said process may be adapted to produce Neu5Acylated i.e. sialylated biomolecules and biomolecules including a saccharide, a peptide, a protein, a glycopeptide, a glycoprotein, a glycolipid, a glycan, an antibody, and a glycoconjugate, in particular, an antibody drug conjugate, and a carbohydrate conjugate vaccine, or a flavonoid.
A method for producing cytidine 5′-monophospho-N-acetyl-neuraminic acid (CMP-Neu5Ac, 1)
The method according to claim 11, wherein the solid support is composed of beads or resins comprising a polymer with epoxide functional groups, with amino epoxide functional groups, with ethylenediamine functional groups, with amino C2 functional groups, with amino C6 functional groups, with anionic/amino C6 spacer functional groups.
A set of enzymes comprising a set of enzymes comprising an N-acylglucosamine 2-epimerase (AGE, EC 5.1.3.8), an N-acetylneuraminate lyase (NAL, EC 4.1.3.3), an N-acylneuraminate cytidylyltransferase (CSS, EC 2.7.7.43), a uridine kinase (UDK, EC 2.7.1.48), a uridine monophosphate kinase (URA6, EC 2.7.4.22) and a polyphosphate kinase 3 (PPK3, EC 2.7.4.1), wherein the set of enzymes is co-immobilized on a polymer by covalent bonds.
Dependencies are inferred from the claim text.