申请人:Max-Planck-Gesellschaft zur Förderung der
Wissenschaften e.V.
公开号:EP3892731A1
公开(公告)日:2021-10-13
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.
本发明涉及一种用低成本底物N-乙酰-D-葡萄糖胺(GlcNAc)、丙酮酸、胞苷和多聚磷酸在单一反应混合物中生产胞苷-5'-单磷酸-N-乙酰-神经氨酸(CMP-Neu5Ac,1)的方法、在单一反应混合物中与一组可选固定化或可选共同固定化的酶进行反应,这些酶包括 N-乙酰葡糖胺 2-酰亚胺酶(AGE)、N-乙酰神经氨酸裂解酶(NAL)、N-乙酰神经氨酸胞苷酸转移酶(CSS)、尿苷激酶(UDK)、单磷酸尿苷激酶和多磷酸激酶 3(PPK3)。此外,所述工艺还可用于生产 Neu5Acylated 即糖醛酸化的生物大分子和生物大分子,包括糖、肽、蛋白质、糖肽、糖蛋白、糖脂、聚糖、抗体和糖共轭物,特别是抗体药物共轭物和碳水化合物共轭疫苗或黄酮类化合物。