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.
- General base
- Glycosyl fluoride
- Inverting glycoside hydrolase
- Reducing-end xylose-releasing exo-oligoxylanase
ASJC Scopus subject areas