Enzymatic route to preparative-scale synthesis of UDP-GlcNAc/GalNAc, their analogues and GDP-fucose
Enzymatic route to preparative-scale synthesis of UDP-GlcNAc/GalNAc, their analogues and GDP-fucose
复制标题
DOI：
10.1038/nprot.2010.3
复制
发表时间：
2010-01-01
期刊：
影响因子：
14.8
通讯作者：
Wang, Peng George
中科院分区：
文献类型：
--
作者：
Zhao, Guohui;Guan, Wanyi;Wang, Peng George
使用糖基转移酶进行酶促合成是高效、高选择性构建多糖的有效方法。糖核苷酸是 Leloir 型糖基转移酶的酶促糖基化反应中的基本供体分子。这些捐助者的申请因其数量有限而受到限制。在此方案中，N-乙酰氨基葡萄糖 (GlcNAc)/N-乙酰半乳糖胺 (GalNAc) 被 N-乙酰己糖胺 1-激酶 (NahK) 磷酸化，随后被 N-乙酰氨基葡萄糖尿苷基转移酶 (GlmU) 焦磷酸化，得到 UDP-GlcNAc/GalNAc。还可以根据这些酶对修饰糖底物的耐受性来制备其他 UDP-GlcNAc/GalNAc 类似物。 GDP-岩藻糖以 L-岩藻糖为原料，由一种双功能酶 L-岩藻糖焦磷酸化酶 (FKP) 通过两个反应构建而成。
Enzymatic synthesis using glycosyltransferases is a powerful approach to building polysaccharides with high efficiency and selectivity. Sugar nucleotides are fundamental donor molecules in enzymatic glycosylation reactions by Leloir-type glycosyltransferases. The applications of these donors are restricted by their limited availability. In this protocol, N-acetylglucosamine (GlcNAc)/N-acetylgalactosamine (GalNAc) are phosphorylated by N-acetylhexosamine 1-kinase (NahK) and subsequently pyrophosphorylated by N-acetylglucosamine uridyltransferase (GlmU) to give UDP-GlcNAc/GalNAc. Other UDP-GlcNAc/GalNAc analogues can also be prepared depending on the tolerance of these enzymes to the modified sugar substrates. Starting from l-fucose, GDP-fucose is constructed by one bifunctional enzyme l-fucose pyrophosphorylase (FKP) via two reactions.