Alternative strategy for converting an inverting glycoside hydrolase into a glycosynthase

Yuji Honda, Shinya Fushinobu, Masafumi Hidaka, Takayoshi Wakagi, Hirofumi Shoun, Hajime Taniguchi, Motomitsu Kitaoka

Research output: Contribution to journalArticlepeer-review

53 Citations (Scopus)

Abstract

The tyrosine residue Y198 is known to support a nucleophilic water molecule with the general base residue, D263, in the reducing-end xylose-releasing exo-oligoxylanase (Rex). A mutation in the tyrosine residue changing it into phenylalanine caused a drastic decrease in the hydrolytic activity and a small increase in the F- releasing activity from α-xylobiosyl fluoride in the presence of xylose. In contrast, mutations at D263 resulted in the decreased F- releasing activity. As a result of the high F- releasing activity and low hydrolytic activity, Y198F of Rex accumulates a large amount of product during the glycosynthase reaction. We propose a novel method for producing a glycosynthase from an inverting glycoside hydrolase by mutating a residue that holds the nucleophilic water molecule with the general base residue while keeping the general base residue intact.

Original languageEnglish
Pages (from-to)325-330
Number of pages6
JournalGlycobiology
Volume18
Issue number4
DOIs
Publication statusPublished - 2008 Apr
Externally publishedYes

Keywords

  • General base
  • Glycosyl fluoride
  • Glycosynthase
  • Inverting glycoside hydrolase
  • Reducing-end xylose-releasing exo-oligoxylanase

ASJC Scopus subject areas

  • Biochemistry

Fingerprint

Dive into the research topics of 'Alternative strategy for converting an inverting glycoside hydrolase into a glycosynthase'. Together they form a unique fingerprint.

Cite this