Abbott D Wade, Boraston Alisdair B
Lethbridge Research Station, Agriculture and Agri-Food Canada, Lethbridge, Alberta, Canada.
Methods Enzymol. 2012;510:211-31. doi: 10.1016/B978-0-12-415931-0.00011-2.
Carbohydrate-binding modules (CBMs) are important components of carbohydrate-active enzymes. Their primary functions are to assist in substrate turnover by targeting appended catalytic modules to substrate and concentrating appended catalytic modules on the surface of substrate. Presented here are four well-established methodologies for investigating and quantifying the CBM-polysaccharide binding relationship. These methods include: (1) the solid state depletion assay, (2) affinity gel electrophoresis, (3) UV difference and fluorescence spectroscopy, and (4) isothermal titration calorimetry. In addition, entropy-driven CBM-crystalline cellulose binding events and differential approaches to calculating stoichiometry with polyvalent polysaccharide ligands are also discussed.
碳水化合物结合模块（CBMs）是碳水化合物活性酶的重要组成部分。它们的主要功能是通过将附加的催化模块靶向底物并将附加的催化模块集中在底物表面来协助底物周转。本文介绍了四种成熟的用于研究和量化CBM-多糖结合关系的方法。这些方法包括：（1）固态消耗测定法，（2）亲和凝胶电泳，（3）紫外差示光谱法和荧光光谱法，以及（4）等温滴定量热法。此外，还讨论了熵驱动的CBM-结晶纤维素结合事件以及用多价多糖配体计算化学计量的差异方法。
