Abstract
Removal of the catalytic nucleophile Glu134 of the retaining 1,3-1,4-β-glucanase from Bacillus licheniformis by mutation to alanine yields an enzyme with no glycosidase activity. The mutant is able to catalyze the regio- and stereospecific glycosylation of α-laminaribiosyl fluoride with different glucoside acceptors through a single-step inverting mechanism. The main advantage of the mutant as glycosylation catalyst with respect to the kinetically controlled transglycosylation using the wild-type enzyme is that the reaction products cannot be hydrolyzed by the mutant enzyme, and glycosylation yields rise to 90%. Copyright (C) 1998 Federation of European Biochemical Societies.
Original language | English |
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Pages (from-to) | 208-212 |
Number of pages | 5 |
Journal | FEBS Letters |
Volume | 440 |
Issue number | 1-2 |
DOIs | |
Publication status | Published - 27 Nov 1998 |
Keywords
- Enzymatic glycosylation
- Glycosyl fluoride
- Glycosynthase
- Nucleophile residue
- β-Glucanase