The Transglycosylation Activity of the Recombinant Endo-.BETA.-N-Acetylglucosaminidase from Mucor hiemalis in Media Containing Organic Solvents

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  • 有機溶媒含有系におけるＭｕｃｏｒ　ｈｉｅｍａｌｉｓ由来リコンビナントエンド‐β‐Ｎ‐アセチルグルコサミニダーゼの糖鎖転移活性
  • ユウキ ヨウバイ ガンユウケイ ニ オケル Mucor hiemalis ユライ リコンビナントエンド ベータ N アセチルグルコサミニダーゼ ノ トウサ テンイ カッセイ ガン エイブン

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Abstract

We investigated the transglycosylation activity of the recombinant endo-β-N-acetylglucosaminidase (Endo-M) from Mucor hiemalis expressed in Candida boidinii in media containing organic solvents, using a disialo biantennary complex-type oligosaccharide from the hen egg yolk glycopeptide as the glycosyl donor and p-nitrophenyl N-acetyl-β-D-glucosaminide as the glycosyl acceptor. The recombinant Endo-M had sufficient transglycosylation activity to transfer the oligosaccharide to pNP-GlcNAc in up to 30-40% organic solvents (v/v) such as acetone, dimethyl sulfoxide and methanol. The transglycosylation activity in each organic solvent differed from each other. The investigation of the donor concentrations in the organic solvents revealed that the increase in the concentrations significantly increased the transglycosylation yields. Using the Endo-M transglycosylation system in organic solvents, we successfully demonstrated the transglycosylation reaction to the glycosyl acceptors which were sparingly soluble in water such as artificial N-acylated D-glucosamines and a 2-O-glycosylated disaccharide.

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