Screening Glycosyltransferases for Polyphenol Modifications

Methods Mol Biol. 2017:1539:229-236. doi: 10.1007/978-1-4939-6691-2_14.

Abstract

Glycosyltransferases offer the opportunity to glycosylate a variety of substrates including health beneficial molecules like flavonoids in a regiospecific manner. Flavonoids are plant secondary metabolites that have antimicrobial, antioxidative, and health beneficial effects. Glycosylation often has impact on these properties and furthermore enhances the water solubility, the stability, and the bioavailability of the molecules. To detect flavonoid glycosylating enzymes we established a metagenome screen for the discovery of modifying clones. This function based screening technique can furthermore detect other modifications like methylations. The method relies on analysis of the culture supernatant extracts from biotransformation reactions in a thin layer chromatography (TLC) approach.

Keywords: Biotransformation; Flavonoid; Glycosyltransferase; META; TLC.

MeSH terms

  • Biotransformation
  • Chromatography, Thin Layer
  • Enzyme Activation
  • Enzyme Assays / methods
  • Gene Library
  • Glycosylation
  • Glycosyltransferases / genetics
  • Glycosyltransferases / metabolism*
  • Metagenome
  • Metagenomics / methods
  • Polyphenols / genetics
  • Polyphenols / metabolism*
  • Substrate Specificity

Substances

  • Polyphenols
  • Glycosyltransferases