One set system for the synthesis and purification of glycosaminoglycan oligosaccharides reconstructed using a hyaluronidase-immobilized column.
One set system for the synthesis and purification of glycosaminoglycan oligosaccharides reconstructed using a hyaluronidase-immobilized column.
复制标题
使用透明质酸酶固定柱重建的一套用于合成和纯化糖胺聚糖寡糖的系统。
作者：
Suto;S.;Kakizaki;I.;Nakamura;T.;and Endo;M.
利用转糖基化反应作为透明质酸酶水解的逆反应，可以按照初步规划的排列方式重构天然糖胺聚糖（GAGs），合成新的人工低聚糖。然而，由于该方法存在反应产物收率低、分离纯化过程复杂等问题，因此研究了该方法的改进方法。转糖基化反应是用装在柱中的牛睾丸透明质酸酶固定树脂进行的。在转糖基化反应中，使用对水解敏感性和转糖基化反应最小尺寸的pyridylaminated (PA) GAG六糖作为受体，而使用大尺寸的GAG作为供体。反应混合物在透明质酸酶固定化树脂柱中孵育后混合，然后引入连续连接的HPLC柱，由三个步骤构建：第一步采用离子交换高效液相色谱法将新合成的GAG寡糖作为反应产物进行浓缩，第二步采用反相高效液相色谱法将PA寡糖与非PA寡糖分离，第三步采用粒度分馏高效液相色谱法对新合成的寡糖进行分离。经过一个完整的转糖基化反应和分离循环，得到了新合成的低聚糖。©2013 Wiley期刊公司生物工程学报（自然科学版），2014。
Using the transglycosylation reaction as a reverse reaction for the hydrolysis of hyaluronidase, new artificial oligosaccharides may be synthesized by reconstructing natural glycosaminoglycans (GAGs) according to preliminary planned arrangements. However, as some problems have been associated with the method, including the low yields of reaction products and complicated processes of separation and purification, improvements in this method were investigated. Transglycosylation reactions were carried out using bovine testicular hyaluronidase‐immobilized resin packed in a column. For the transglycosylation reaction, pyridylaminated (PA) GAG hexasaccharides, which were the minimum size for hydrolysis sensitivity and the transglycosylation reaction, were used as acceptors, whereas large size GAGs were used as donors. The reaction mixture was pooled after incubation in the hyaluronidase‐immobilized resin column and was then introduced into continuously joined HPLC columns constructed from three steps: the first step of ion‐exchange HPLC for concentrating newly synthesized GAG oligosaccharides as reaction products, the second step of reverse phase HPLC for separating PA oligosaccharides from non‐PA oligosaccharides, and the third step of size fractionation HPLC for fractionating newly synthesized oligosaccharides. Newly synthesized oligosaccharides were obtained by one complete cycle of the transglycosylation reaction and separation. © 2013 Wiley Periodicals, Inc. Biopolymers 101: 189–196, 2014.