Tanaka Hidenori, Yoshimura Yayoi, Hindsgaul Ole
Carlsberg Laboratory, Copenhagen, Denmark.
Curr Protoc Nucleic Acid Chem. 2013 Oct 8;54:13.12.1-13.12.10. doi: 10.1002/0471142700.nc1312s54.
Chemoenzymatic oligosaccharide synthesis is attractive since it eliminates the tedious multistep protection-deprotection requirements of pure chemical synthesis. Chemoenzymatic synthesis using glycosyltransferases, however, requires not only the correct enzyme to control both regio- and stereospecificity, but also the glycosyl donor to provide the sugar that is added. This unit describes a simple synthesis of sugar-nucleoside diphosphates (sugar-NDPs), the type of glycosyl donor (e.g., UDP-Glc, UDP-Gal, ADP-Glc) required by most glycosyltransferases, by using a chemical coupling reaction in water. The preparation of sugar-NDPs by this method therefore does not require any skills in synthetic organic chemistry.
化学酶促寡糖合成很有吸引力，因为它省去了纯化学合成中繁琐的多步保护-脱保护步骤。然而，使用糖基转移酶进行化学酶促合成不仅需要合适的酶来控制区域和立体特异性，还需要糖基供体来提供添加的糖。本单元描述了一种通过在水中进行化学偶联反应来简单合成糖核苷二磷酸（糖-NDPs）的方法，糖-NDPs是大多数糖基转移酶所需的糖基供体类型（例如，UDP-Glc、UDP-Gal、ADP-Glc）。因此，用这种方法制备糖-NDPs不需要任何有机合成化学技能。
