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dc.contributor.authorChoe, J.H.
dc.contributor.authorVanderNoot, V.A.
dc.contributor.authorLinhardt, Robert J.
dc.contributor.authorDordick, J.S.
dc.datePolysaccharide lyases that can degrade glycosaminoglycans (GAGs) were identified in an anaerobic strain living in the human intestine. The strain was isolated from the stool of a healthy male and identified as Bacteroides sp. strain HJ-15. A detailed taxonomical study indicated the species is a strain of Bacteroides stercoris. The isolate was cultured and the polysaccharide lyase activity was partially purified. This enzyme preparation could act on GAGs containing either glucosamine or galactosamine suggesting the presence of both heparinases and chondroitinases. Various GAGs were incubated with the partially purified enzyme and the products formed were analyzed by strong anion-exchange high performance liquid chromatography and proton nuclear magnetic resonance spectroscopy. These studies demonstrated the presence of at least two types of polysaccharide lyases: heparin lyase and chondroitin sulfate lyase. The eliminative mechanism of these lyase enzymes was confirmed through the isolation of unsaturated disaccharide products. The heparin lyase acted on both heparin and acharan sulfate, a GAG recently isolated from Achatina fulica. The Bacteroides chondroitin lyase, acted on chondroitin sulfates A, B (dermatan sulfate), and C, resembling chondroitin lyase ABC. The presence of a GAG-degrading organism in human intestine may pose problems for the effective oral administration of GAG drugs.
dc.dateDes polysaccharides lyases capables de d6grader des glycosaminoglycanes (GAGs) ont ete identifiees dans une souche anaerobie vivant dans l'intestin humain. La souche a 6t6 isolee a partir d'excrements d'homme sain et identi iee a Bacteroides sp. HJ-15. Une etude taxonomique d6taill6e a permis d'etablir que ces especes appartenaient A la souche Bacteroides stercoris. Apres isolement, cette souche a ete cultiv6e et l'activite polysaccharide lyase a 6t6 partiellement purifiee. La preparation enzymatique etait active sur des GAGs contenant des glucosamines ou galactosamines, sugg6rant la presence d'h6parinases et de chondroitinases. Differents GAGs ont ete incub6s en presence de l'enzyme partiellement purifiee et les produits formes analyses par chromatographie liquide echangeuse d'anions et par spectroscopie de resonance magnetique nucleaire du proton. Ces etudes ont demontre la presence d'au moins deux types de polysaccharides lyases, soit l'hoparine lyase et la chondroitine sulfate lyase. Le mecanisme d'elimination de ces enzymes lyases a ete confirme par isolement des produits disaccharides insatures. L'heparine lyase etait active sur 1'heparine et l'acharane sulfate, un GAG recemment isole d'Achatina fulica. La chondroitine lyase de Bacteroides fut active sur les chondroitines sulfates A, B (dermatane sulfate) et C. La presence d'un organisme degradant les GAGs dans l'intestin humain est susceptible de poser un probleme pour l'administration orale efficace de medicaments a base de GAGs.
dc.date.accessioned2022-06-27T16:21:51Z
dc.date.available2022-06-27T16:21:51Z
dc.date.issued1998
dc.identifier.citationResolution of Structurally Similar Glycoproteins by Affinity-BasedReversed Micellar Extraction and Separation, J.-H. Choe, V.A. VanderNoot,R.J. Linhardt, J.S. Dordick, AIChE Journal, 44, 2542-2548, 1998.
dc.identifier.urihttps://doi.org/10.1002/aic.690441121
dc.identifier.urihttps://hdl.handle.net/20.500.13015/5890
dc.descriptionAIChE Journal, 44, 2542-2548
dc.descriptionNote : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
dc.description.abstractAffinity-based reversed micellar extraction and separation (ARMES) has proven effective in separating glycoproteins from nonglycosylated proteins from natural sources. The ability of ARMES to resolve closely related glycoproteins is of paramount importance if ARMES is to be used in glycoform resolution. It is demonstrated that ARMES can resolve the structurally similar soybean peroxidase (SBP; MW 37 kDa, pI 4.1) and αt-acid glycoprotein (AGP; MW 43 kDa, pI 3.7), both of which have affinity for Concanavalin A (Con A) (the affinity ligand). SBP was almost exclusively extracted at pH 8 and above, with a separation factor greater than 50 (resolution ∼ 20), far better than was possible using Con A affinity chromatography (R ∼ 0.25, separation factor ∼ 2). Model calculations suggest that differences in affinity measured by an equilibrium-building assay cannot account for the favorable extraction of SBP over AGP at higher pH. Hydrophobic interactions and/or charge shielding appear to affect partitioning of the lectin - glycoprotein complexes and add greatly to the selectivity of extraction in ARMES, especially at higher pH values.
dc.description.urihttps://login.libproxy.rpi.edu/login?url=https://doi.org/10.1002/aic.690441121
dc.languageen_US
dc.language.isoENG
dc.relation.ispartofThe Linhardt Research Labs Online Collection
dc.relation.ispartofRensselaer Polytechnic Institute, Troy, NY
dc.relation.urihttps://harc.rpi.edu/
dc.subjectBiology
dc.subjectChemistry and chemical biology
dc.subjectChemical and biological engineering
dc.subjectBiomedical engineering
dc.titleResolution of Structurally Similar Glycoproteins by Affinity-BasedReversed Micellar Extraction and Separation
dc.typeArticle
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dc.creator.identifierhttps://orcid.org/0000-0003-2219-5833
dc.relation.departmentThe Linhardt Research Labs.
dc.relation.departmentThe Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)


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