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通过部分木葡聚糖酶消化对木葡聚糖寡糖进行测序:两种新四糖的制备及其定量和定性分析。

Sequencing of xyloglucan oligosaccharides by partial Driselase digestion: the preparation and quantitative and qualitative analysis of two new tetrasaccharides.

作者信息

Lorences E P, Fry S C

机构信息

Division of Biological Sciences, University of Edinburgh, United Kingdom.

出版信息

Carbohydr Res. 1994 Oct 17;263(2):285-93. doi: 10.1016/0008-6215(94)00170-7.

Abstract

The pentasaccharide (XXG), [formula: see text] obtained from Rosa xyloglucan, was converted to two isomeric tetrasaccharides, a and b (Xyl1.Glc3), by mild acid hydrolysis. During hydrolysis in 2 M trifluoroacetic acid at 90 degrees C, optimal yields of a and b were obtained after 20-40 min. Each tetrasaccharide was purified by preparative paper chromatography and high-pressure liquid chromatography (HPLC). The two isomers were distinguished by the products of their partial digestion with Driselase, which hydrolyses the glucosidic bonds sequentially from the non-reducing terminus: a and b yielded cellobiose and Xyl-->Glc-->Glc, respectively showing that they were [formula: see text] and [formula: see text] respectively. Tetrasaccharide b was chromatographically identical, upon HPLC on Dionex CarboPac PA1, with the tetrasaccharide produced from XXG by the action of Tropaeolum alpha-D-xylosidase, supporting the proposed structure. Xyloglucan oligosaccharides were assayed quantitatively by measurement of the yield of isoprimeverose (Xyl-->Glc) after complete Driselase digestion.

摘要

从罗莎木葡聚糖中获得的五糖(XXG),[化学式:见原文],通过温和酸水解转化为两种异构四糖,a和b(木糖1.葡萄糖3)。在90℃的2M三氟乙酸中水解时,20 - 40分钟后获得a和b的最佳产率。每种四糖通过制备性纸色谱法和高压液相色谱法(HPLC)进行纯化。这两种异构体通过用Driselase部分消化的产物来区分,Driselase从非还原末端依次水解糖苷键:a和b分别产生纤维二糖和木糖→葡萄糖→葡萄糖,表明它们分别是[化学式:见原文]和[化学式:见原文]。在Dionex CarboPac PA1上进行HPLC时,四糖b在色谱上与通过旱金莲α - D - 木糖苷酶作用从XXG产生的四糖相同,支持了所提出的结构。通过测量完全Driselase消化后异麦芽糖(木糖→葡萄糖)的产率对木葡聚糖寡糖进行定量分析。

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