Morell M K, Samuel M S, O'Shea M G
CSIRO Plant Industry, Camberra, ACT, Australia.
Electrophoresis. 1998 Nov;19(15):2603-11. doi: 10.1002/elps.1150191507.
The analysis of the fine structure of starches is important to the investigation of linkages between starch structure and function and to the investigation of the properties and roles of starch biosynthetic, modifying and degradation enzymes. Fluorophore-assisted carbohydrate electrophoresis has recently been introduced as a method for the analysis of the oligosaccharide populations released by the enzymatic digestion of starches, which has advantages in resolution and sensitivity over previously used methods, and provides the capacity for the facile analysis of oligosaccharide populations on either a molar or mass basis. The use of fluorophore-assisted carbohydrate electrophoresis for the analysis of oligosaccharides is reviewed with particular reference to the choice of label, efficiency of labeling and separation techniques. Examples of separations using slab gel electrophoresis, DNA sequencer analysis and capillary electrophoresis are presented and we conclude that on the basis of resolution and reproducibility, capillary electrophoresis is the method of choice for the separation of oligosaccharides of degree of polymerization from 1 to 100. Examples of isoamylase-debranched starches and glycogens analyzed by capillary electrophoresis are presented. The capillary electrophoresis analysis of starch structure through the analysis of oligosaccharides released by the debranching of limit dextrins derived from starches and glycogens is introduced as a useful diagnostic of starch structure. The potential for future development of novel diagnostics for starch structure using fluorophore-assisted carbohydrate electrophoresis is discussed.
淀粉精细结构的分析对于研究淀粉结构与功能之间的联系以及淀粉生物合成、修饰和降解酶的性质与作用至关重要。荧光团辅助碳水化合物电泳最近被引入作为分析淀粉酶解释放的寡糖群体的方法，它在分辨率和灵敏度方面优于先前使用的方法，并能够在摩尔或质量基础上轻松分析寡糖群体。本文综述了荧光团辅助碳水化合物电泳在寡糖分析中的应用，特别提及了标记物的选择、标记效率和分离技术。给出了使用平板凝胶电泳、DNA测序仪分析和毛细管电泳进行分离的实例，我们得出结论，基于分辨率和重现性，毛细管电泳是分离聚合度为1至100的寡糖的首选方法。展示了通过毛细管电泳分析异淀粉酶脱支淀粉和糖原的实例。介绍了通过分析淀粉和糖原衍生的极限糊精脱支释放的寡糖来进行淀粉结构的毛细管电泳分析，这是一种有用的淀粉结构诊断方法。讨论了使用荧光团辅助碳水化合物电泳开发新型淀粉结构诊断方法的未来潜力。
