The α-Thioglycoligase Derived from a GH89 α-N-Acetylglucosaminidase Synthesises α-N-Acetylglucosamine-Based Glycosides of Biomedical Interest
详细信息 查看全文
- 作者：Ndivhuwo Olga Tshililo ; Andrea Strazzulli ; Beatrice Cobucci-Ponzano ; Luisa Maurelli ; Roberta Iacono ; Emiliano Bedini ; Maria Michela Corsaro ; Erick Strauss and Marco Moracci
- 刊名：Advanced Synthesis & Catalysis
- 出版年：2017
- 出版时间：February 20, 2017
- 年：2017
- 卷：359
- 期：4
- 页码：663-676
- 全文大小：2450K
- ISSN：1615-4169
文摘
We report here on the preparation of a novel α-thioglycoligase that can be used for the fast and efficient synthesis of α-N-acetylglucosamine-based glycosides. Using the α-N-acetyl-glucosaminidase from Clostridium perfringens of family GH89 (according to the Carbohydrate Active Enzymes classification) as starting point, we prepared mutants in the acid/base residue glutamic acid 483 that were found to have different synthetic efficiencies (maximal yields >80% after 24 h) in the presence of an activated donor and suitable acceptors. The synthetic potential of the Glu483 alanine mutant as an α-thioglycoligase – only the third biocatalyst with this stereospecificity reported to date, and the first selective for the N-acetylglucosamine moiety – was demonstrated by producing for the first time N-acetyl-α-d-glucosaminyl azide and N-acetylglucosamine α-thioglycosides in high yields. To showcase the application of such compounds, we show that they offer the wild-type CpGH89 protection from thermal unfolding, demonstrating their potential as pharmacological chaperones for the treatment of mucopolysaccharidosis IIIB (Sanfilippo syndrome).