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An efficient chemoenzymatic strategy for synthesizing complex glycosphingolipids by enzymatic extension of lactosyl sphingosine (LacβSph, 3) using sequential one-pot multienzyme (OPME) reactions with C18-cartridge purification after each glycosylation reaction followed by a simple acylation process. The structures of target α-Gal pentasaccharyl sphingosine Galα3nLc4βSph (2) and α-Gal pentasaccharyl ceramide Galα3nLc4βCer (1) are also shown.
