2. Academic Validation
  3. Simultaneous one-pot multienzymatic synthesis and purification of Uridine-5'-Diphosphate-α-D-glucuronide and Uridine-5'-Diphosphate-α-d-xylose

Simultaneous one-pot multienzymatic synthesis and purification of Uridine-5'-Diphosphate-α-D-glucuronide and Uridine-5'-Diphosphate-α-d-xylose

  • Carbohydr Res. 2025 Aug:554:109516. doi: 10.1016/j.carres.2025.109516.
Arnaud Pâris 1,  Tiffany Rundstadler 1,  Pierre Lafite 2
Affiliations

Affiliations

  • 1 Institut de Chimie Organique et Analytique (ICOA), UMR 7311, CNRS-Université d'Orléans, Université d'Orléans, BP 6759, Orléans, 45067 Cedex 2, France.
  • 2 Institut de Chimie Organique et Analytique (ICOA), UMR 7311, CNRS-Université d'Orléans, Université d'Orléans, BP 6759, Orléans, 45067 Cedex 2, France. Electronic address: [email protected].
PMID: 40383045 DOI: 10.1016/j.carres.2025.109516
Abstract

Glucuronosyltransferases and xylosyltransferases are glycosyltransferases (GT) that are involved in a wide range of biological processes, including xenobiotic metabolism or proteoglycan biosynthesis and require as substrates activated sugar donors, respectively UDP-α-D-glucuronide (UDP-GlcA) and UDP-α-d-xylose (UDP-Xyl). As both nucleotide sugars are hardly available, we have developed a one-pot multienzymatic synthesis to produce sequentially UDP-GlcA and UDP-Xyl. This methodology involves the production of UDP-GlcA for glucuronic acid, coupling glucuronokinase and UDP-sugar pyrophosphorylase activities, followed by UDP-GlcA decarboxylation by UDP-Xylose Synthase. Each of these UDP-sugars can then be simultaneously purified and isolated from the unique reaction mixture, providing an efficient production methodology of these rare UDP-sugars for subsequent enzymatic studies of corresponding GT.

Keywords

Biocatalysis; Glycosyltransferase; Nucleotide-sugars; UDP-Sugar pyrophosphorylase; UDP-Xylose synthase.

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