Preparative and Kinetic Analysis of β-1,4- and β-1,3-Glucan Phosphorylases Informs Access to Human Milk Oligosaccharide Fragments and Analogues Thereof

Research output: Contribution to journalArticlepeer-review

  • External authors:
  • Ravindra Pal Singh
  • Giulia Pergolizzi
  • Sergey A Nepogodiev
  • Peterson de Andrade
  • Sakonwan Kuhaudomlarp

Abstract

The enzymatic synthesis of oligosaccharides depends on the availability of suitable enzymes, which remains a limitation. Without recourse to enzyme engineering or evolution approaches, herein we demonstrate the ability of wild-type cellodextrin phosphorylase (CDP: β-1,4-glucan linkage-dependent) and laminaridextrin phosphorylase (Pro_7066: β-1,3-glucan linkage-dependent) to tolerate a number of sugar-1- phosphate substrates, albeit with reduced kinetic efficiency. In spite of catalytic efficiencies of <1 % of the natural reactions, we demonstrate the utility of given phosphorylase-sugar phosphate pairs to access new-to-nature fragments of human milk oligosaccharides, or analogues thereof, in multi-milligram quantities.

Bibliographical metadata

Original languageEnglish
Pages (from-to)1043-1049
Number of pages7
JournalCHEMBIOCHEM
Volume21
Issue number7
Early online date30 Dec 2019
DOIs
Publication statusPublished - 1 Apr 2020