Highly efficient transglycosylation of sialo-complex-type oligosaccharide using Coprinopsis cinerea endoglycosidase and sugar oxazoline
Highly efficient transglycosylation of sialo-complex-type oligosaccharide using Coprinopsis cinerea endoglycosidase and sugar oxazoline
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DOI：
10.1007/s10529-016-2230-0
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发表时间：
2017-01-01
影响因子：
2.7
通讯作者：
Takegawa, Kaoru
中科院分区：
文献类型：
--
作者：
Higuchi, Yujiro;Eshima, Yasunari;Takegawa, Kaoru
为了建立一种有效的化学酶修饰方法，使糖蛋白的N-连接寡糖链结构均一，这是影响其生物活性的关键因素。pH 7.5对于SG-恶唑啉的稳定性和Endo-CC的转糖基化反应都是理想的。用于生产糖基化RNA酶B的最有效的反应条件是：80 μ g GlcNAc-RNase B，200 μ g SG-恶唑啉和3 μ g Endo-CCN 180 H在20 μ l 20 mM Tris/HCl pH 7.5中于30 ℃下反应30-60分钟。CCN 180 H有利于生产用均质双触角复合型寡糖修饰的药物糖蛋白。
To establish an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities.Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-beta-N-acetylglucosaminidase (Endo-CCN180H) were employed. pH 7.5 was ideal for both SG-oxazoline's stability and Endo-CC's transglycosylation reaction. The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 mu g GlcNAc-RNase B, 200 mu g SG-oxazoline and 3 mu g Endo-CCN180H in 20 mu l 20 mM Tris/HCl pH 7.5 at 30 A degrees C for 30-60 min.This transglycosylation method using SG-oxazoline and Endo-CCN180H is beneficial for producing pharmaceutical glycoproteins modified with homogenous biantennary-complex-type oligosaccharides.