4-Thiocellooligosaccharides. Their synthesis and use as ligands for the separation of cellobiohydrolases of Trichoderma reesei by affinity chromatography.
4-Aminophenyl 1,4-dithio-beta-cellobioside (6) was obtained by treatment of methyl 2,3,6-tri-O-benzoyl-4-O-triflyl-alpha-D-galactopyranoside with the sodium salt of 1-thio-beta-D-glucopyranose, followed by acetolysis and glycosylation of the corresponding bromide with 4-aminobenzenethiol and subsequent deacylation. A similar synthesis starting with the 1-thiolate of 1,4-dithio-beta-cellobiose led to the trisaccharide 4-aminophenyl 1,4,4'-trithiocellotrioside (16). The 4-acetamidophenyl di- and tri-thiocellooligosaccharides were found to be excellent competitive inhibitors of the hydrolysis of 4-methylumbelliferyl beta-lactoside with respective Ki values of 25 and 6.5 mM. The two 4-aminophenyl oligosaccharides 6 and 16 were coupled to CH-Sepharose 4B, and the affinity gels were used for the purification of cellobiohydrolases from a crude commercial cellulolytic extract of T. reesei. Cellobiohydrolases I or II were selectively desorbed from gels bearing ligands 6 and 16.
Orgeret
C
C
Centre de Recherches sur les Macromolécules Végétales, C.N.R.S., Grenoble, France.
Seillier
E
E
Gautier
C
C
Defaye
J
J
Driguez
H
H
D002240:Carbohydrate Sequence; D002475:Cellobiose / Q000031:analogs & derivatives*; D043366:Cellulose 1,4-beta-Cellobiosidase; D002846:Chromatography, Affinity / Q000379:methods*; D006026:Glycoside Hydrolases / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D013865:Thioglycosides / Q000138:chemical synthesis / Q000378:metabolism*; D014242:Trichoderma / Q000201:enzymology*
D013865:Thioglycosides; D002475:Cellobiose; D006026:Glycoside Hydrolases; D043366:Cellulose 1,4-beta-Cellobiosidase