Valent B S, Darvill A G, McNeil M, Robertsen B K, Albersheim P
Carbohydr Res. 1980 Mar;79(2):165-92. doi: 10.1016/s0008-6215(00)83830-6.
This paper describes a new glycosyl-sequencing method. This method was made possible by the ability to fractionate complex mixtures of peralkylated oligosaccharides by reversed-phase, high-pressure liquid chromatography. The fractionation ability of the reversed-phase system allows the isolation and subsequent unambiguous identification by g.l.c.-m.s. of disaccharides, almost all trisaccharides, and, in some cases, tetrasaccharides generated by successive partial acid hydrolysis, reduction, and ethylation of a permethylated, complex carbohydrate. As these small oligosaccharides overlap within the unhydrolyzed, complex carbohydrate, the oligosaccharide sequences may be pieced together, and, with the glycosyl-linkage composition of the intact complex carbohydrate, can be used to determine the glycosyl sequence of the complex carbohydrate. The details of the sequencing method are illustrated by the elucidation of the glycosyl sequences of three complex carbohydrates. These examples demonstrate the wide variety of complex carbohydrates whose structures can be ascertained by the new sequencing technique. Two of the examples are the commercially available polysaccharides, lichenan and xanthan, whose structures have already been reported. The other example is a nonasaccharide derived from xyloglucan, a structural polymer of plant cell-walls. The glycosyl residues of the complex carbohydrates studied include hexosyl, deoxyhexosyl, pentosyl, glycosyluronic, and pyruvic acetal-substituted hexosyl residues. It will be demonstrated that the new glycosyl-sequencing technique is not compromised by the presence, in the carbohydrate to be analyzed, of glycosyl linkages possessing very different acid labilities. Two major advantages of this sequencing technique are that it is relatively rapid and that it requires only milligram quantities of carbohydrate.
本文描述了一种新的糖基测序方法。这种方法得以实现是因为能够通过反相高压液相色谱法对全烷基化寡糖的复杂混合物进行分馏。反相系统的分馏能力允许通过气相色谱 - 质谱法分离并随后明确鉴定由全甲基化的复杂碳水化合物连续部分酸水解、还原和乙基化产生的二糖、几乎所有的三糖以及某些情况下的四糖。由于这些小寡糖在未水解的复杂碳水化合物中相互重叠，寡糖序列可以拼凑起来，并结合完整复杂碳水化合物的糖基连接组成，用于确定复杂碳水化合物的糖基序列。通过阐明三种复杂碳水化合物的糖基序列来说明测序方法的细节。这些例子展示了可以通过这种新测序技术确定其结构的多种复杂碳水化合物。其中两个例子是市售多糖，地衣聚糖和黄原胶，其结构已经有报道。另一个例子是源自木葡聚糖（植物细胞壁的一种结构聚合物）的九糖。所研究的复杂碳水化合物的糖基残基包括己糖基、脱氧己糖基、戊糖基、糖醛酸基和丙酮酸缩醛取代的己糖基残基。将证明，新的糖基测序技术不会因待分析碳水化合物中存在酸稳定性差异很大的糖基连接而受到影响。这种测序技术的两个主要优点是相对快速，并且只需要毫克量的碳水化合物。
