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- Title
Highly efficient transglycosylation of sialo-complex-type oligosaccharide using Coprinopsis cinerea endoglycosidase and sugar oxazoline.
- Authors
Higuchi, Yujiro; Eshima, Yasunari; Huang, Yibo; Kinoshita, Takashi; Sumiyoshi, Wataru; Nakakita, Shin-ichi; Takegawa, Kaoru
- Abstract
Objectives: To establish an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities. Results: Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β- N-acetylglucosaminidase (Endo-CC) 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 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CC in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. Conclusions: This transglycosylation method using SG-oxazoline and Endo-CC is beneficial for producing pharmaceutical glycoproteins modified with homogenous biantennary-complex-type oligosaccharides.
- Subjects
OLIGOSACCHARIDE biotechnology; GLYCOSYLATION; ENDOGLYCOSIDASES; COMPOSITION of mushrooms; OXAZOLINE; GLYCOPROTEINS
- Publication
Biotechnology Letters, 2017, Vol 39, Issue 1, p157
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-016-2230-0