Sequential One-Pot Multienzyme Chemoenzymatic Synthesis of Glycosphingolipid Glycans

Abstract

Glycosphingolipids are a diverse family of biologically important glycolipids. In addition to variations on the lipid component, more than 300 glycosphingolipid glycans have been characterized. These glycans are directly involved in various molecular recognition events. Several naturally occurring sialic acid forms have been found in sialic acid-containing glycosphingolipids, namely gangliosides. However, ganglioside glycans containing less common sialic acid forms are currently not available. Herein, highly effective one-pot multienzyme (OPME) systems are used in sequential for high-yield and cost-effective production of glycosphingolipid glycans, including those containing different sialic acid forms such as N-acetylneuraminic acid (Neu5Ac), N-glycolylneuraminic acid (Neu5Gc), 2-keto-3-deoxy-D-glycero-D-galacto-nononic acid (Kdn), and 8-O-methyl-N-acetylneuraminic acid (Neu5Ac8OMe). A library of 64 structurally distinct glycosphingolipid glycans belonging to ganglio-series, lacto-/neolacto-series, and globo-/isoglobo-series glycosphingolipid glycans is constructed. These glycans are essential standards and invaluable probes for bioassays and biomedical studies.

Machine‐Driven Enzymatic Oligosaccharide Synthesis by Using a Peptide Synthesizer

“…Ganglioside GM1 is a glycan epitope located on the cell surface and plays important roles in many biological processes such as virus-cell interactions. [24] The enzymatic synthesis of GM1 from lactose involves three glycosylation steps. [24] The enzymes used are Pasteurella multocida α2,3-sialyltransferase (PmsT1), Campylobacter jejuni β1,4-N-acetylgalactosaminyltransferase (CgtA) and Campylobacter jejuni β1,3-galactosyltransferase CgtB.…”

“…[24] The enzymatic synthesis of GM1 from lactose involves three glycosylation steps. [24] The enzymes used are Pasteurella multocida α2,3-sialyltransferase (PmsT1), Campylobacter jejuni β1,4-N-acetylgalactosaminyltransferase (CgtA) and Campylobacter jejuni β1,3-galactosyltransferase CgtB. [24] The standardized procedure contains three processes (Figure 4B): automated synthesis, product release, and product purification.…”

“…[24] The enzymes used are Pasteurella multocida α2,3-sialyltransferase (PmsT1), Campylobacter jejuni β1,4-N-acetylgalactosaminyltransferase (CgtA) and Campylobacter jejuni β1,3-galactosyltransferase CgtB. [24] The standardized procedure contains three processes (Figure 4B): automated synthesis, product release, and product purification. In order to adapt the CEM peptide synthesizer for this purpose, several optimization procedures were required.…”

Enzymatic and Chemoenzymatic Syntheses of Disialyl Glycans and Their Necrotizing Enterocolitis Preventing Effects

“…The relatively low yield was due to GalβSEt ( 17 ) being a less optimal acceptor than GalNAcβOR in GM2 or GD2 ganglioside oligosaccharides. 34 …”

Chemoenzymatic Synthesis of DSGb5 and Sialylated Globo‐series Glycans

“…[6] By treating NK cells with sialidase to remove NeuAc and disrupt the cis interactions,t he unmasked Siglec-7 can interact with DSGb5-Cer on the surface of the human renal cancer cell line ACHN,decreasing the cytotoxicity of the NK cells and promoting the metastasis and invasion of RCC. [8] Obtaining as ufficient amount of pure DSGb5 is key to elucidating its mechanism of action and to further develop cancer immunotherapies.C larifying the binding activities of sialylated globo-series glycans and the effect of the NeuAc position on their interactions with Siglec-7 is also of great interest.Thee nzymatic and chemoenzymatic syntheses of globoseries glycans,such as Gb5, SSEA-4, and Globo H, have been reported by several research groups, [9][10][11] but no method has been reported for the synthesis of DSGb5. DSGb5 has aunique glycan structure with two NeuAcs as a2,3-and a2,6linkages on the nonreducing-end galactose (Gal) and Nacetylgalactosamine (GalNAc), respectively,o fG b5 (Figure 1).…”