Controllable Enzymatic Synthesis of Natural Asymmetric Human Milk Oligosaccharides DOI Creative Commons

Hsien‐Wei Tseng,

Hsin‐Kai Tseng,

Kai-Eng Ooi

et al.

JACS Au, Journal Year: 2024, Volume and Issue: 4(11), P. 4496 - 4506

Published: Nov. 2, 2024

Among human milk oligosaccharides (HMOs), linear HMOs are synthesized through mature but varied routes. Although branched can be by chemical, enzymatic, or chemoenzymatic methods, these methods cannot easily applied to the synthesis of asymmetric multiantennary oligosaccharides. Herein, we developed a controllable method synthesize biantennary HMOs. In our synthetic route, GlcNAcβ1,3(GlcN3β1,6)Glaβ1,4Glc was first chemically as core tetrasaccharide, which contains β1,6GlcN3 "stop" sugar in transferase-catalyzed glycosylation. The desired sugars at GlcNAcβ1–3Gal arm assembled using galactosyltransferase, N-acetylglucosaminyltransferase, and fucosyltransferase. Then, Staudinger reduction acetylation were used transform GlcN3 GlcNAc assemble initiating "go" process. By manipulating glycosylations, 22 natural synthesized.

Language: Английский

EASyMap-Guided Stepwise One-Pot Multienzyme (StOPMe) Synthesis and Multiplex Assays Identify Functional Tetraose-Core-Human Milk Oligosaccharides DOI Creative Commons
Yuanyuan Bai, Anand K. Agrahari, Libo Zhang

et al.

JACS Au, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 27, 2025

Carbohydrates are biologically and medicinally important molecules that attracting growing attention to their synthesis applications. Unlike the biosynthetic processes for nucleic acids proteins, carbohydrate biosynthesis is not template-driven, more challenging, often leads product variations. In lieu of templates biosynthesis, we describe herein a new concept designing enzyme assembly synthetic maps (EASyMaps) as blueprints guide glycosyltransferase-dependent stepwise one-pot multienzyme (StOPMe) systematically access structurally diverse carbohydrates in target-oriented manner. The strategy demonstrated construction comprehensive library tetraose-core-containing human milk oligosaccharides (HMOs) presenting functional glycan epitopes shared by complex HMOs. tetraose-core-HMOs attractive candidates large-scale production development HMO-based nutraceuticals. To achieve preparative-scale targets containing Neu5Acα2–6GlcNAc component, α2–6-sialyltransferase hST6GALNAC5 successfully expressed E. coli. Neoglycoproteins with controlled valencies prepared immobilized on fluorescent magnetic beads. Multiplex bead assays reveal ligands glycan-binding proteins from plants, influenza viruses, human, bacteria, identifying promising HMO EASyMaps StOPMe systematic manner broadly applicable beyond efficient process suitable can be potentially adapted automation.

Language: Английский

Citations

1
Host–Guest Chemistry-Mediated Biomimetic Chemoenzymatic Synthesis of Complex Glycosphingolipids DOI

Yuan Ma,

Yating Liu, Chang Cao

et al.

Journal of the American Chemical Society, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 11, 2025

Glycosphingolipids (GSLs) are amphipathic complex biomolecules constituted of hydrophilic glycans covalently linked to hydrophobic lipids via glycosidic bonds. GSLs widely distributed in cells and tissues, where they play crucial roles various biological functions disease processes. However, the heterogeneity complexity make it difficult explore their precise biofunctions due obstacles obtaining well-defined structures. Herein, we report a host–guest-chemistry-mediated biomimetic chemoenzymatic approach for efficient synthesis diverse GSLs. A key feature this is that use methyl-β-cyclodextrin enables glycolipids forming water-soluble inclusion complexes improve solubility aqueous media, thereby facilitating enzyme-catalyzed reactions. The power applicability our demonstrated by streamlined biologically important globo-, ganglio-, neolacto-, lacto-series library containing 20 neutral acidic with different fucosylation sialylation patterns. developed method will open new avenues easily access wide range biomedical applications.

Language: Английский

Citations

1
Ketodeoxynonulosonic Acid Hydroxylase (Kdnase) Assisted Site‐Specific Enzymatic α2,6‐Sialylation DOI Open Access

Yu Zhou,

Yun Li, Jiayu Wen

et al.

Chinese Journal of Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 26, 2025

Comprehensive Summary Owing to its promiscuous substrate specificity and high catalytic efficiency, the bacterial α2,6‐sialyltransferase from Photobacterium damselae (Pd2,6ST) has been widely used for synthesis of various α2,6‐linked sialosides. However, Pd2,6ST is not a suitable enzyme regioselective α2,6‐sialylation complex acceptor substrates containing multiple galactose (Gal) and/or N ‐acetylgalactosamine (GalNAc) residues due specificity. In this study, novel enzymatic engineering strategy was developed overcome limitation by employing enzymatically introduced ketodeoxynonulosonic acid (Kdn) as temporary “protecting group” at unwanted sialylation sites. The Kdn can be selectively removed hydrolase Aspergillus fumigatus ( Af Kdnase) appropriate stage without affecting coexisting sialic residues, such ‐acetylneuraminic (Neu5Ac) or ‐glycolylneuraminic (Neu5Gc). This provides general practical approach sialosides, including sialylated poly‐LacNAc glycans, disialylated ganglioside glycan epitopes, branched human milk oligosaccharides.

Language: Английский

Citations

0
Controllable Enzymatic Synthesis of Natural Asymmetric Human Milk Oligosaccharides DOI Creative Commons

Hsien‐Wei Tseng,

Hsin‐Kai Tseng,

Kai-Eng Ooi

et al.

JACS Au, Journal Year: 2024, Volume and Issue: 4(11), P. 4496 - 4506

Published: Nov. 2, 2024

Among human milk oligosaccharides (HMOs), linear HMOs are synthesized through mature but varied routes. Although branched can be by chemical, enzymatic, or chemoenzymatic methods, these methods cannot easily applied to the synthesis of asymmetric multiantennary oligosaccharides. Herein, we developed a controllable method synthesize biantennary HMOs. In our synthetic route, GlcNAcβ1,3(GlcN3β1,6)Glaβ1,4Glc was first chemically as core tetrasaccharide, which contains β1,6GlcN3 "stop" sugar in transferase-catalyzed glycosylation. The desired sugars at GlcNAcβ1–3Gal arm assembled using galactosyltransferase, N-acetylglucosaminyltransferase, and fucosyltransferase. Then, Staudinger reduction acetylation were used transform GlcN3 GlcNAc assemble initiating "go" process. By manipulating glycosylations, 22 natural synthesized.

Language: Английский

Citations

3