Efficient chemoenzymatic synthesis of an N-glycan isomer library

Lei Li, Yunpeng Liu, Cheng Ma, Jingyao Qu, Angie D. Calderon, Baolin Wu, Na Wei, Xuan Wang, Yuxi Guo, Zhongying Xiao, Jing Song, Go Sugiarto, Yanhong Li, Hai Yu, Xi Chen, Peng George Wang

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59 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)5652-5661
Number of pages10
JournalChemical Science
Volume6
Issue number10
DOIs
StatePublished - Jun 22 2015

ASJC Scopus subject areas

  • Chemistry(all)

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    Li, L., Liu, Y., Ma, C., Qu, J., Calderon, A. D., Wu, B., Wei, N., Wang, X., Guo, Y., Xiao, Z., Song, J., Sugiarto, G., Li, Y., Yu, H., Chen, X., & Wang, P. G. (2015). Efficient chemoenzymatic synthesis of an N-glycan isomer library. Chemical Science, 6(10), 5652-5661. https://doi.org/10.1039/c5sc02025e