RESUMEN
The mono-glucose-branched cyclodextrins having an appropriate spacer between the beta-cyclodextrin and a glucose moiety were synthesized from beta-cyclodextrin and arbutin. They had the significantly high association constants for doxorubicin, the anticancer agent, in the range of 10(5)-10(6)M(-1), and worked as highly reactive glycosyl acceptors for the transglycosylation reaction by endo-beta-N-acetylglucosaminidase of Mucor hiemalis to produce sialo-complex type oligosaccharide-branched cyclodextrins in the high yields of 65-67%.
Asunto(s)
Acetilglucosaminidasa/metabolismo , Ciclodextrinas/síntesis química , Doxorrubicina/química , Portadores de Fármacos/síntesis química , Mucor/enzimología , Antineoplásicos/administración & dosificación , Sitios de Unión , Ciclodextrinas/química , Sistemas de Liberación de Medicamentos , Glucosa/química , Glicosilación , Modelos Moleculares , Oligosacáridos/químicaRESUMEN
We found that the recombinant endo-beta-N-acetylglucosaminidase of Mucor hiemalis (Endo-M) expressed in Candida boidinii had the transglycosylation activity of transferring a bisecting hybrid-type oligosaccharide from an ovalbumin glycopeptide to the acceptor (p-nitrophenyl 2-acetamido-2-deoxy-beta-D-glucopyranoside) in a good yield of 43%.
Asunto(s)
Acetilglucosamina/análogos & derivados , Acetilglucosaminidasa/metabolismo , Glicopéptidos/metabolismo , Mucor/enzimología , Oligosacáridos/metabolismo , Acetilglucosamina/metabolismo , Clonación Molecular , Glicosilación , Ovalbúmina , Proteínas RecombinantesRESUMEN
We investigated the transglycosylation reaction of the recombinant endo-beta-N-acetylglucosaminidase from Mucor hiemalis (Endo-M) expressed in Candida boidinii using such sugar derivatives as N-acylated d-glucosamines, C-glucosyl derivatives, and a 2-O-glycosylated disaccharide as acceptors. We found that a variety of sugar derivatives modified at C-1 or C-2 could be used as acceptors for transglycosylation by Endo-M to create novel oligosaccharides.
Asunto(s)
Manosil-Glicoproteína Endo-beta-N-Acetilglucosaminidasa/química , Mucor/enzimología , Oligosacáridos/química , Acetilglucosamina/análogos & derivados , Acetilglucosamina/química , Disacáridos/química , Glicosilación , Estructura MolecularRESUMEN
We examined the transglycosylation reaction by the recombinant endo-beta-N-acetylglucosaminidase from Mucor hiemalis (Endo-M) expressed in Candida boidinii in media containing organic solvents. The recombinant Endo-M could transglycosylate a disialo biantennary complex-type oligosaccharide from hen egg yolk glycopeptide to p-nitrophenyl N-acetyl-beta-D-glucosaminide even in the presence of 30% acetone, dimethyl sulfoxide, or methanol. The yield of the transglycosylation product reached 21-34% of the total amount of acceptor, while the yield was only about 14% in aqueous solution.