Family 1 glycosyltransferases (GT1, UGTs) are subject to dilution-induced inactivation and low chemo stability toward their own acceptor substrates

David Teze*, Gonzalo Nahuel Bidart, Ditte Hededam Welner

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

Glycosylation reactions are essential but challenging from a conventional chemistry standpoint. Conversely, they are biotechnologically feasible as glycosyltransferases can transfer sugar to an acceptor with perfect regio- and stereo-selectivity, quantitative yields, in a single reaction and under mild conditions. Low stability is often alleged to be a limitation to the biotechnological application of glycosyltransferases. Here we show that these enzymes are not necessarily intrinsically unstable, but that they present both dilution-induced inactivation and low chemostability towards their own acceptor substrates, and that these two phenomena are synergistic. We assessed 18 distinct GT1 enzymes against three unrelated acceptors (apigenin, resveratrol, and scopoletin—respectively a flavone, a stilbene, and a coumarin), resulting in a total of 54 enzymes: substrate pairs. For each pair, we varied catalyst and acceptor concentrations to obtain 16 different reaction conditions. Fifteen of the assayed enzymes (83%) displayed both low chemostability against at least one of the assayed acceptors at submillimolar concentrations, and dilution-induced inactivation. Furthermore, sensitivity to reaction conditions seems to be related to the thermal stability of the enzymes, the three unaffected enzymes having melting temperatures above 55°C, whereas the full enzyme panel ranged from 37.4 to 61.7°C. These results are important for GT1 understanding and engineering, as well as for discovery efforts and biotechnological use.

Original languageEnglish
Article number909659
JournalFrontiers in Molecular Biosciences
Volume9
Number of pages8
ISSN2296-889X
DOIs
Publication statusPublished - 2022

Bibliographical note

Funding Information:
The authors thank the Novo Nordisk Foundation for financial support (grants NNF18OC0034744, NNF10CC1016517, NNF19OC0055620, and NNF20CC0035580).

Publisher Copyright:
Copyright © 2022 Teze, Bidart and Welner.

Keywords

  • biotechnology
  • glycosylation
  • glycosyltransferases
  • GT1
  • polyphenols
  • stability
  • UGT

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