Quantification, characterization and biofunctional studies of N-glycans on proteins remain challenging tasks due to the complexity, diversity and low abundance of these glycans. The availability of structurally defined N-glycan (especially isomer) libraries is essential to help solve these tasks. We report herein an efficient chemoenzymatic strategy, namely Core Synthesis/Enzymatic Extension (CSEE), for rapid production of diverse N-glycans. Starting with 5 chemically prepared building blocks, 8 N-glycan core structures containing one or two terminal N-acetyl-D-glucosamine (GlcNAc) residue(s) were chemically synthesized via consistent use of oligosaccharyl thioethers as glycosylation donors in a convergent fragment coupling strategy. Each of these core structures was then extended to 5 to 15 N-glycan sequences by enzymatic reactions catalyzed by 4 robust glycosyltransferases. Success in synthesizing N-glycans with Neu5Gc and core-fucosylation further expanded the ability of the enzymatic extension. Meanwhile, high performance liquid chromatography with an amide column enabled rapid and efficient purification (>98% purity) of N-glycans in milligram scales. A total of 73 N-glycans (63 isomers) were successfully prepared and characterized by MS2 and NMR. In summary, the CSEE strategy provides a practical approach for “mass production” of structurally defined N-glycans, which are important standards and probes for glycoscience.
Li, Lei; Liu, Yunpeng; Ma, Cheng; Qu, Jingyao; Calderon, Angie D.; Wu, Baolin; Wei, Na; Wang, Xuan; Guo, Yuxi; Xiao, Zhongying; Song, Jing; Sugiarto, Go; Li, Yanhong; Yu, Hai; Chen, Xi; and Wang, Peng George, "Efficient Chemoenzymatic Synthesis of an N-glycan Isomer Library" (2015). Chemistry Faculty Publications. 12.
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Originally Published in:
Chem Sci, 6 (10), 5652-5661. DOI: 10.1039/c5sc02025e