@article {1436,
	title = {Glycosynthase-based synthesis of xylo-oligosaccharides using an engineered retaining xylanase from Cellulomonas fimi},
	journal = {Organic \& Biomolecular Chemistry},
	volume = {4},
	number = {10},
	year = {2006},
	note = {ISI Document Delivery No.: 041NHTimes Cited: 20Cited Reference Count: 39},
	pages = {2025-2032},
	type = {Article},
	abstract = {Glycosynthases are synthetic enzymes derived from retaining glycosidases in which the catalytic nucleophile has been replaced. The mutation allows irreversible glycosylation of sugar acceptors using glycosyl fluoride donors to afford oligosaccharides without any enzymatic hydrolysis. Glycosynthase technology has proven fruitful for the facile synthesis of useful oligosaccharides, therefore the expansion of the glycosynthase repertoire is of the utmost importance. Herein, we describe for the first time a glycosynthase, derived from a retaining xylanase, that synthesizes a range of xylo-oligosaccharides. The catalytic domain of the retaining endo-1,4-beta-xylanase from Cellulomonas fimi (CFXcd) was successfully converted to the corresponding glycosynthase by mutation of the catalytic nucleophile to a glycine residue. The mutant enzyme (CFXcd-E235G) was found to catalyze the transfer of a xylobiosyl moiety from alpha-xylobiosyl fluoride to either p-nitrophenyl beta-xylobioside or benzylthio beta-xylobioside to afford oligosaccharides ranging in length from tetra- to dodecasaccharides. These products were purified by high performance liquid chromatography in greater than 60\% combined yield. H-1 and C-13 NMR spectroscopic analyses of the isolated p-nitrophenyl xylotetraoside and p-nitrophenyl xylohexaoside revealed that CFXcd-E235G catalyzes both the regio- and stereo-selective synthesis of xylo-oligosaccharides containing, exclusively, beta-(1 {\textendash}> 4) linkages.},
	keywords = {4-GLYCANASE, BETA-1, DIRECTED EVOLUTION, EFFICIENT SYNTHESIS, ENZYMATIC-SYNTHESIS, GLYCOSIDASES, MASS-SPECTROMETRY, SPECIFICITY, SUBSTRATE, TRANSGLYCOSYLATION, XYLOOLIGOSACCHARIDES},
	isbn = {1477-0520},
	url = {<Go to ISI>://000237462400022},
	author = {Kim, Y. W. and Fox, D. T. and Hekmat, O. and Kantner, T. and McIntosh, L. P. and Warren, R. A. J. and Withers, S. G.}
}
@article {3114,
	title = {IN-VIVO INHIBITION OF BETA-GLUCOSIDASE AND BETA-MANNOSIDASE ACTIVITY IN RATS BY 2-DEOXY-2-FLUORO-BETA-GLYCOSYL FLUORIDES AND RECOVERY OF ACTIVITY IN-VIVO AND IN-VITRO},
	journal = {Biochemical Journal},
	volume = {301},
	year = {1994},
	note = {ISI Document Delivery No.: PA559Times Cited: 12Cited Reference Count: 25Part 2},
	month = {Jul},
	pages = {343-348},
	type = {Article},
	abstract = {2-Deoxy-2-fluoro-beta-glucosyl and -beta-mannosyl fluorides administered to rats in a single dose(10 mg/kg) inhibited beta-glucosidase and beta-mannosidase activity respectively after 1 h in brain, spleen, liver and kidney tissues. This inhibition, presumably caused by accumulation of 2-deoxy-2-fluoroglycosyl-enzyme intermediates, indicates that intact 2-deoxy-2-fluoroglycosyl fluorides are distributed to these organs and, in the case of brain, that they cross the blood/brain barrier. beta-Glucosidase activity recovered completely or partially in brain, spleen, liver and kidney by 20-48 h. beta-Mannosidase activity partially recovered in all tissues by 48 h. beta-Galactosidase activity in brain and kidney was not significantly affected by administration of either the gluco or manno compounds at this dosage, indicating that these inhibitors are directed towards specific glycosidases. Observation of similar relatively rapid rates of beta-glycosidase re-activation in vivo and in tissue homogenates in vitro at 37 degrees C suggests that hydrolysis or transglycosylation of 2-deoxy-2-fluoroglycosyl-enzymes, not protein synthesis, are the primary mechanisms involved in the recovery of glycosidase activity inhibited by this class of compounds in vivo.},
	keywords = {DEFICIENCY, ENZYME INTERMEDIATE, FORMS, GLUCOCEREBROSIDASE, GLYCOSIDASES, INACTIVATION, LIVER, MICE},
	isbn = {0264-6021},
	url = {<Go to ISI>://A1994PA55900006},
	author = {McCarter, J. D. and Adam,Michael J. and Hartman, N. G. and Withers, S. G.}
}
