Fucosylated carbohydrate determinants are common components of cell surface glycoconjugates and secreted unconjugated glycans, which play pivotal roles in many physiological and pathological processes. The biosynthesis of Lewis antigens involves multiple fucosyltransferases that catalyse the fucosylation of the poly-N-acetyllactosamine carbohydrate backbone in a non-site-specific manner and thus generate heterogeneous and incompletely fucosylated Lewis antigen regioisomers. In this study, an α2,6-sialylation module was used to introduce α2,6-linked sialic acid to specific sites as the protecting group against fucosylation, thus precisely controlling enzymatic fucosylation of poly-N-acetyllactosamine glycans in a site-specific manner. The sialic acid protecting group can be easily removed by sialidase after fucosylation to provide a variety of fucosylated poly-N-acetyllactosamine glycans with defined fucosylation patterns. The general applicability and robustness of this reprogrammed enzymatic assembly line was exemplified in the synthesis of 22 complex Lewis antigens and chimeric histo-blood group antigens with a total of 10 enzyme modules for the construction of 10 different glycosidic linkages.
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The NMR spectroscopy, LC-MS, MS2 and other data that support the findings of this study are available from the corresponding author upon reasonable request.
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This project was financially supported by the National Natural Science Foundation of China (Grant nos. 21672128, 21877072 and 21807064), State Key Laboratory of Microbial Technology (M2016-06), Department of Science and Technology of Shandong Province (2016GSF121002, ZR201709190252 and 2016GGH4502) and Shandong University (2018JC053). The authors thank X. Chen at the University of California, Davis and X. Huang at Michigan State University for reading the manuscript and helpful discussion.
The authors declare no competing interests.
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Ye, J., Xia, H., Sun, N. et al. Reprogramming the enzymatic assembly line for site-specific fucosylation. Nat Catal 2, 514–522 (2019). https://doi.org/10.1038/s41929-019-0281-z
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