Highly efficient synthesis of beta(1 -> 4)-oligo- and -polysaccharides using a mutant cellulase

Citation
S. Fort et al., Highly efficient synthesis of beta(1 -> 4)-oligo- and -polysaccharides using a mutant cellulase, J AM CHEM S, 122(23), 2000, pp. 5429-5437
Citations number
54
Categorie Soggetti
Chemistry & Analysis",Chemistry
Journal title
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
ISSN journal
00027863 → ACNP
Volume
122
Issue
23
Year of publication
2000
Pages
5429 - 5437
Database
ISI
SICI code
0002-7863(20000614)122:23<5429:HESOB->2.0.ZU;2-#
Abstract
This report describes an efficient chemoenzymatic synthesis of a variety of regioselectively modified beta(1-->4)-oligo- and -polysaccharides. This su ccessfull approach was based on: (i) the use of a "glycosynthase" which is a Glu-197-Ala nucleophile mutant of the retaining cellulase endoglucanase I (Cel7B) from Humicola insolens and (ii) the rational design of modified ac ceptor and donor molecules through a careful examination of information giv en by the X-ray structures of wild type and mutated enzymes. The mutant was able to catalyze, in high yield, the regio- and stereoselective glycosylat ion of alpha-glycobiosyl fluorides both unsubstituted and modified with var ious mono- and disaccharide accepters, as well as the polymerization of the se donors through a single-step inverting mechanism.