Department of Chemistry, University of California, Davis, California.
Curr Protoc. 2021 Mar;1(3):e91. doi: 10.1002/cpz1.91.
Gangliosides are biologically important sialic acid-containing glycolipids found commonly in human and other vertebrates. Isolation of pure gangliosides from cells or tissues is difficult, and chemical synthesis of gangliosides usually involves numerous steps with low synthetic yields. We report here a chemoenzymatic synthesis and purification protocol for two ganglioside cancer antigens, GM3 and GD3. One-pot multienzyme glycosylation reactions are used to sequentially prepare GM3 and GD3 sphingosines from chemically synthesized lactosyl sphingosine. A facile C18-cartridge purification procedure after each glycosylation reaction provides the desired pure glycosyl sphingosine product, which is readily acylated to form the target ganglioside. © 2021 Wiley Periodicals LLC. Basic Protocol: Chemoenzymatic synthesis and purification of GM3 and GD3 gangliosides Support Protocol: Monitoring reactions by thin-layer chromatography.
神经节苷脂是一类生物学上重要的含唾液酸的糖脂，在人和其他脊椎动物中普遍存在。从细胞或组织中分离出纯的神经节苷脂很困难，而神经节苷脂的化学合成通常需要经过多步反应，且产率较低。本文报告了两种神经节苷脂癌症抗原 GM3 和 GD3 的化学酶法合成和纯化方案。一锅多酶糖基化反应用于从化学合成的乳糖基神经酰胺中依次制备 GM3 和 GD3 的神经酰胺。在每次糖基化反应后，采用简单的 C18 柱纯化程序，可提供所需的纯糖苷神经酰胺产物，该产物可轻松酰化形成目标神经节苷脂。© 2021 Wiley Periodicals LLC. 基本方案：GM3 和 GD3 神经节苷脂的化学酶法合成和纯化 支持方案：通过薄层层析监测反应。
