Biochemical and in silico characterization of glycosyltransferases from red sweet cherry (Prunus avium L.) reveals their broad specificity toward phenolic substrates

Daniel Clayton-Cuch; Long Yu; Daniel McDougal; Crista A Burbidge; John B Bruning; David Bradley; Christine Böttcher; Vincent Bulone

doi:10.1016/j.fochms.2023.100193

Biochemical and in silico characterization of glycosyltransferases from red sweet cherry (Prunus avium L.) reveals their broad specificity toward phenolic substrates

Food Chem (Oxf). 2023 Dec 31:8:100193. doi: 10.1016/j.fochms.2023.100193. eCollection 2024 Jul 30.

Authors

Daniel Clayton-Cuch^{1

2}, Long Yu¹, Daniel McDougal³, Crista A Burbidge², John B Bruning³, David Bradley⁴, Christine Böttcher², Vincent Bulone^{1

5}

Affiliations

¹ Adelaide Glycomics, University of Adelaide, School of Agriculture, Food and Wine, Waite Campus, Adelaide, South Australia 5064, Australia.
² CSIRO, Waite Campus, Glen Osmond, South Australia 5064, Australia.
³ Institute for Photonics and Advanced Sensing (IPAS), School of Biological Sciences, The University of Adelaide, Adelaide, South Australia 5005, Australia.
⁴ Agilent Technologies Australia Pty Ltd, Mulgrave, Melbourne, Victoria 3171, Australia.
⁵ Division of Glycoscience, Department of Chemistry, School of Engineering Sciences in Chemistry, Biotechnology and Health, Royal Institute of Technology (KTH), AlbaNova University Centre, Stockholm 10691, Sweden.

Abstract

Polyphenolic compounds are a class of phytonutrients that play important roles in plants and contribute to human health when incorporated into our diet through fruit consumption. A large proportion occur as glycoconjugates but the enzymes responsible for their glycosylation are poorly characterized. Here, we report the biochemical and structural characterization of two glycosyltransferases from sweet cherry named PaUGT1 and PaUGT2. Both are promiscuous glucosyltransferases active on diverse anthocyanidins and flavonols, as well as phenolic acids in the case of PaUGT1. They also exhibit weaker galactosyltransferase activity. The expression of the gene encoding PaUGT1, the most active of the two proteins, follows anthocyanin accumulation during fruit ripening, suggesting that this enzyme is the primary glycosyltransferase involved in flavonoid glycosylation in sweet cherry. It can potentially be used to synthesize diverse glycoconjugates of flavonoids for integration into bioactive formulations, and for generating new fruit cultivars with enhanced health-promoting properties using breeding methods.

Keywords: Anthocyanins; Cherry; Flavonols; Glycosyltransferase; Phenolic compounds; Protein structure modelling.