Enzymatic Synthesis of Glycosides and Oligosaccharides

Enzymatic synthesis of glycosides and oligosaccharides harnesses nature’s catalysts to assemble complex carbohydrates with high regio- and stereoselectivity under mild conditions. Glycoside phosphorylases and related glycosyltransferases achieve bond formation by transferring sugar moieties from activated donors—often sugar phosphates or nucleotide sugars—onto diverse acceptors, ranging from simple alcohols to elaborate oligosaccharide chains. This biocatalytic approach offers distinct advantages over purely chemical methods, including reduced reaction steps, lower energy input and diminished reliance on protecting-group chemistry. Engineering of active-site residues and whole-enzyme scaffolds has expanded substrate ranges and enhanced stability, enabling tailor-made routes to bioactive oligosaccharides, prebiotic fibres and glycosylated natural products. Structural studies, kinetic analysis and mechanistic insights have deepened understanding of sugar-binding modes, catalytic loops and domain architecture, guiding the rational design of enzymes for industrial bioprocesses. The global significance of these developments spans pharmaceuticals, functional foods, cosmetics and biomaterials, where precisely defined glycosides and oligosaccharides fulfil roles as stabilisers, immunomodulators and scaffolds for advanced therapeutics.

Research from Nature Portfolio

Recent studies have elucidated structural bases for substrate specificity in glycoside phosphorylases. One investigation of a β-1,2-oligoglucan phosphorylase revealed a unique N-terminal β-sandwich domain that positions oligosaccharide substrates between this domain and the catalytic core. Distinctive flipping of the sugar ring at the +1 subsite induces bond distortion that fine-tunes selectivity for longer β-1,2-glucooligosaccharides, offering a blueprint for engineering new linkage specificities. Another study characterised a thermostable cellodextrin phosphorylase from a deep-sea thermophile, uncovering dual cellobiose and cellodextrin activity. This enzyme displays optimal function at elevated temperature and accepts substrates of varying chain length, enabling efficient synthesis of cellodextrins with degrees of polymerisation above two. The combination of broad substrate scope, high catalytic efficiency and inherent thermostability highlights its promise as a robust catalyst for large-scale oligosaccharide manufacture.

Enzymatic Synthesis of Glycosides and Oligosaccharides publication trend

The graph below shows the total number of articles in enzymatic synthesis of glycosides and oligosaccharides across all publications each year (not limited to Nature Index journals).

Technical terms

Glycoside: A molecule in which a sugar is bound to another functional group via a glycosidic bond.

Oligosaccharide: A carbohydrate composed of a small number (typically three to ten) of monosaccharide units linked by glycosidic bonds.

Glycoside phosphorylase: An enzyme that catalyses reversible phosphorolysis of glycosidic bonds, forming sugar 1-phosphates and shorter sugars.

Phosphorolysis: A reaction in which a glycosidic bond is cleaved by inorganic phosphate, yielding a sugar phosphate and a free sugar.

Transglycosylation: The transfer of a sugar moiety from one molecule (donor) to another (acceptor) without free phosphate release, often catalysed by hydrolases under specific conditions.

Donor substrate: The activated sugar molecule (e.g., sugar phosphate or nucleotide sugar) that provides the glycosyl unit in enzymatic synthesis.

Acceptor substrate: The molecule (e.g., alcohol, sugar, polyol) that receives the glycosyl unit during glycoside formation.

References

1. Sucrose Phosphorylase and Related Enzymes in Glycoside Hydrolase Family 13: Discovery, Application and Engineering. International Journal of Molecular Sciences (2020).
2. β-Glucan phosphorylases in carbohydrate synthesis. Applied Microbiology and Biotechnology (2021).
3. Enzymatic synthesis using glycoside phosphorylases. Carbohydrate Research (2014).
4. Identification of Euglena gracilis β-1,3-glucan phosphorylase and establishment of a new glycoside hydrolase (GH) family GH149. Journal of Biological Chemistry (2018).
5. Mechanistic insight into the substrate specificity of 1,2-β-oligoglucan phosphorylase from Lachnoclostridium phytofermentans. Scientific Reports (2017).
6. Biochemical properties of GH94 cellodextrin phosphorylase THA_1941 from a thermophilic eubacterium Thermosipho africanus TCF52B with cellobiose phosphorylase activity. Scientific Reports (2017).
7. Whole cell-based catalyst for enzymatic production of the osmolyte 2-O-α-glucosylglycerol. Microbial Cell Factories (2021).