Enzymatic Synthesis of 2-(β-Galactosyl)-ethyl Methacrylate by β-Galactosidase from Pyrococcus woesei and Application for Glycopolymer Synthesis and Lectin Studies

Marius Hoffmann, Elisabeth Gau, Susanne Braun, Andrij Pich*, Lothar Elling*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

9 Citations (Web of Science)

Abstract

Glycosidases have long been used for the synthesis of glycosides by transglycosylation reactions. Especially glycosidases from hyperthermophilic bacteria are useful for reactions under extreme reaction conditions, e.g., in the presence of organic solvents. We herein report the facile enzymatic synthesis and purification of 2-(beta-galactosyl)-ethyl methacrylate (Gal-EMA) with the recombinant hyperthermostable glycosidase from Pyrococcus woesei in high yields. Optimized reaction conditions resulted in gram-scale synthesis of the galactosylated monomer with 88% transglycosylation yield. The product Gal-EMA was characterized by high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS), nuclear magnetic resonance (NMR) spectroscopy, and infrared (IR) spectroscopy. Gal-EMA was utilized to synthesize sugar-functionalized acrylate polymers with defined amounts of incorporated galactose (0-100%). Analysis of the binding affinity of the lectin RCA(120) from Ricinus communis to the glycopolymers using an enzyme-linked lectin assay (ELLA) revealed K-D values between 0.24 and 6.2 nM, depending on the amount of incorporated Gal-EMA. The potential of Gal-EMA for the synthesis of acrylate-functionalized glycan oligomers was demonstrated by sequential elongation of the terminal galactose by two glycosyltransferases, resulting in the terminal glycan N-acetyllactosamine (LacNAc) epitope. In conclusion, the enzymatic synthesis of Gal-EMA opens new routes to a series of novel monomeric building blocks for the synthesis of glycan-functionalized polyacrylates.

Original languageEnglish
Pages (from-to)974-987
Number of pages14
JournalBiomacromolecules
Volume21
Issue number2
DOIs
Publication statusPublished - Feb 2020

Keywords

  • RICINUS-COMMUNIS AGGLUTININ
  • WELL-DEFINED GLYCOPOLYMERS
  • CATALYZED SYNTHESIS
  • CHEMOENZYMATIC SYNTHESIS
  • ESCHERICHIA-COLI
  • BINDING
  • TOXIN
  • POLYMERIZATION
  • GLUCOSIDASE
  • COPOLYMERS

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