Alternative strategy for converting an inverting glycoside hydrolase into a glycosynthase.
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Honda Y, Fushinobu S, Hidaka M, Wakagi T, Shoun H, Taniguchi H, Kitaoka M
Alternative strategy for converting an inverting glycoside hydrolase into a glycosynthase.
Glycobiology. 2008 Apr;18(4):325-30. doi: 10.1093/glycob/cwn011. Epub 2008 Feb 9.
- PubMed ID
- 18263897 [ View in PubMed]
- 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 alpha-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.