Glycosynthase-Mediated Synthesis of Glycosphingolipids

• Published in: Journal of the American Chemical Society Vol. 128; no. 19; pp. 6300 - 6301
• Main Authors: Vaughan, Mark D, Johnson, Karl, DeFrees, Shawn, Tang, Xiaoping, Warren, R. Antony J, Withers, Stephen G
• Format: Journal Article
• Language: English
• Published: WASHINGTON American Chemical Society 17.05.2006; Amer Chemical Soc
• Subjects: Catalysis; Chemistry; Chemistry, Multidisciplinary; Exact sciences and technology; Glucose - analogs & derivatives; Glucose - chemistry; Glycoside Hydrolases - genetics; Glycoside Hydrolases - metabolism; Glycosphingolipids - chemical synthesis; Lipids; Mutation; Organic chemistry; Physical Sciences; Preparations and properties; Science & Technology; Sphingosine - chemistry; GM1 GANGLIOSIDE; ENDOGLYCOCERAMIDASE; ANALOGS; DISEASE; GD3; OLIGOSACCHARIDE; STRAIN; Organic synthesis; Catalytic reaction; Enzyme; Glycosidases; Hydrolases; Enzymatic catalysis; Endoglycosylceramidase; O-Glycosidases; Catalyst activity
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/ja058469n

Glycosphingolipids play crucial roles in virtually every stage of the cell cycle, and their clinical administration has been proposed as a treatment for Alzheimer's, Parkinson's, stroke, and a range of other conditions. However, lack of supply has severely hindered testing of this potential. A novel glycosynthase-based synthetic strategy is demonstrated, involving a mutant of an endoglycoceramidase in which the catalytic nucleophile has been ablated. This mutant efficiently couples a range of glycosyl fluoride donors with a range of sphingosine-based acceptors in yields around 95%. This technology opens the door to large-scale production of glycosphingolipids and, thus, to clinical testing.