Robb Mélissa, Hobbs Joanne K, Boraston Alisdair B
Department of Biochemistry and Microbiology, University of Victoria, 3055, STN CSC, Victoria, BC, Canada, V8W 3P6.
Methods Mol Biol. 2017;1588:215-221. doi: 10.1007/978-1-4939-6899-2_17.
Fluorophore-assisted carbohydrate electrophoresis (FACE) is a method in which a fluorophore is covalently attached to the reducing end of carbohydrates, thereby allowing visualization following high-resolution separation by electrophoresis. This method can be used for carbohydrate profiling and sequencing, as well as for the determination of the specificity of carbohydrate-active enzymes. Here, we describe and demonstrate the use of FACE to separate and visualize the glycans released following digestion of oligosaccharides by glycoside hydrolases (GHs) using two examples: (1) the digestion of chitobiose by the streptococcal β-hexosaminidase GH20C, and (2) the digestion of glycogen by the GH13 member SpuA.
荧光团辅助碳水化合物电泳（FACE）是一种将荧光团共价连接到碳水化合物还原端的方法，从而能够在通过电泳进行高分辨率分离后实现可视化。该方法可用于碳水化合物谱分析和测序，以及确定碳水化合物活性酶的特异性。在此，我们通过两个例子描述并展示了使用FACE来分离和可视化糖苷水解酶（GHs）消化寡糖后释放的聚糖：（1）链球菌β-己糖胺酶GH20C对壳二糖的消化，以及（2）GH13成员SpuA对糖原的消化。
