@article { ISI:000228450200004,
	title = {Engineering of a thioglycoligase: randomized mutagenesis of the acid-base residue leads to the identification of improved catalysts},
	journal = {PROTEIN ENGINEERING DESIGN \& SELECTION},
	volume = {18},
	number = {1},
	year = {2005},
	month = {JAN},
	pages = {33-40},
	abstract = {Thioglycoligases are recently introduced variants of retaining glycosidases in which the acid-base catalyst has been mutated, rendering them capable of thioglycoside synthesis. The original acid-base mutant of Agrobacterium sp. beta-glucosidase (E170A) was previously shown to be an effective thioglycoligase carrying out glycosyltransfer from 2,4-dinitrophenyl glycosides to several different thio sugar acceptors. Here we report the generation of a screen for improved thioglycoligases, randomized mutagenesis of the acid-base catalyst E170 and identification of variants superior to E170A. Furthermore we have established a coupled assay allowing kinetic analysis of isolated variants and found that Abg E170Q is 5-fold faster than Abg E170A when 2,4-dinitrophenyl glucoside is used as donor and 100-fold faster when glucosyl azide is used. To demonstrate its utility, different acceptor and donor sugar combinations were employed to produce thio-linked di- or trisaccharides in high yields, showing the considerable versatility of the system for the synthesis of carbohydrate mimetics.},
	issn = {1741-0126},
	doi = {10.1093/protein/gzi003},
	author = {Mullegger, J and Jahn, M and Chen, HM and WARREN, RAJ and Withers, SG}
}
