Engineering and kinetic characterisation of two glucosyltransferases from Arabidopsis thaliana

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Abstract

This study describes the characterisation of a chimeric mutant derived from two arabidopsis glucosyltransferases, 71C1 and 71C3. A chimera, N1C3, was constructed to contain the N-terminal domain of 71C1 and the C-terminal domain of 71C3. The chimera and the wild-type GTs displayed a similar K-m towards the acceptor scopoletin. However, N1C3 had a K-m near identical to 71C3 towards UDP-glucose, but was three-fold lower than that of 71C1. The results suggest that the acceptor and sugar donor are recognised independently by the N- and C-terminal domain of the GTs respectively, and provide a foundation for the future design of glucosyltransferase biocatalysts through assembling domains with different affinity towards the acceptor and donor. (C) 2008 Elsevier Masson SAS. All rights reserved.

Original languageEnglish
Pages (from-to)830-834
Number of pages5
JournalBIOCHIMIE
Volume90
Issue number5
DOIs
Publication statusPublished - May 2008

Keywords

  • protein engineering
  • glucosyltransferases
  • domain swapping
  • glycosylation
  • enzyme kinetics
  • ENZYME-CATALYZED REACTIONS
  • GLUTATHIONE-S-TRANSFERASE
  • CRYSTAL-STRUCTURE
  • BIOSYNTHETIC-PATHWAY
  • MEDICAGO-TRUNCATULA
  • MULTIGENE FAMILY
  • SMALL MOLECULES
  • GLYCOSYLTRANSFERASES
  • CLASSIFICATION
  • SIMILARITIES

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