Galactosyl-biomimetic dye-ligands for the purification of Dactylium dendroides galactose oxidase

dc.contributor.author	Mazitsos, CF	en
dc.contributor.author	Rigden, DJ	en
dc.contributor.author	Tsoungas, PG	en
dc.contributor.author	Clonis, YD	en
dc.date.accessioned	2014-06-06T06:45:03Z
dc.date.available	2014-06-06T06:45:03Z
dc.date.issued	2002	en
dc.identifier.issn	00219673	en
dc.identifier.uri	http://dx.doi.org/10.1016/S0021-9673(02)00176-0	en
dc.identifier.uri	http://62.217.125.90/xmlui/handle/123456789/2224
dc.subject	Affinity adsorbents	en
dc.subject	Biomimetic ligands	en
dc.subject	Dactylium dendroides	en
dc.subject	Molecular modelling	en
dc.subject	Stationary phases, LC	en
dc.subject.other	Adsorption	en
dc.subject.other	Biomimetic materials	en
dc.subject.other	Dissociation	en
dc.subject.other	Electrophoresis	en
dc.subject.other	Enzymes	en
dc.subject.other	Purification	en
dc.subject.other	Affinity chromatography	en
dc.subject.other	Ligands	en
dc.subject.other	Chromatographic analysis	en
dc.subject.other	adsorbent	en
dc.subject.other	agarose	en
dc.subject.other	alcohol dehydrogenase	en
dc.subject.other	cibacron blue f3ga	en
dc.subject.other	dye	en
dc.subject.other	galactosamine	en
dc.subject.other	galactose oxidase	en
dc.subject.other	glucose dehydrogenase	en
dc.subject.other	glucose oxidase	en
dc.subject.other	ligand	en
dc.subject.other	affinity chromatography	en
dc.subject.other	analytic method	en
dc.subject.other	article	en
dc.subject.other	biomimetics	en
dc.subject.other	controlled study	en
dc.subject.other	dissociation constant	en
dc.subject.other	enzyme activity	en
dc.subject.other	enzyme purification	en
dc.subject.other	gel electrophoresis	en
dc.subject.other	molecular model	en
dc.subject.other	nonhuman	en
dc.subject.other	priority journal	en
dc.subject.other	quantitative analysis	en
dc.subject.other	synthesis	en
dc.subject.other	Chromatography, Liquid	en
dc.subject.other	Coloring Agents	en
dc.subject.other	Electrophoresis, Polyacrylamide Gel	en
dc.subject.other	Galactose	en
dc.subject.other	Galactose Oxidase	en
dc.subject.other	Ligands	en
dc.subject.other	Mass Spectrometry	en
dc.subject.other	Mitosporic Fungi	en
dc.subject.other	Molecular Mimicry	en
dc.subject.other	Nuclear Magnetic Resonance, Biomolecular	en
dc.subject.other	Dendroides	en
dc.subject.other	Hypomyces rosellus	en
dc.title	Galactosyl-biomimetic dye-ligands for the purification of Dactylium dendroides galactose oxidase	en
heal.type	journalArticle	en
heal.identifier.primary	10.1016/S0021-9673(02)00176-0	en
heal.publicationDate	2002	en
heal.abstract	Two anthraquinone galactosyl-biomimetic dye-ligands comprising, as terminal biomimetic moiety, galactose analogues (1-amino-1-deoxy-β-D-galactose and D(+)-galactosamine) were designed for the enzyme galactose oxidase (GAO), using molecular modelling, synthesized and characterized. The biomimetic ligands were immobilized on agarose beads and the affinity adsorbents, together with a non-biomimetic adsorbent bearing Cibacron Blue 3GA, were studied for their ability to purify GAO from Dactylium dendroides. Both biomimetic adsorbents showed higher purifying ability for GAO compared to the non-biomimetic adsorbent, thus demonstrating their superior effectiveness as affinity chromatography materials. In particular, the affinity adsorbent comprising, as terminal biomimetic moiety, 1-amino-1-deoxy-β-D-galactose (BM1) exhibited the highest purifying ability for GAO. This affinity adsorbent did not bind galactose dehydrogenase, glucose dehydrogenase, alcohol dehydrogenase, or glucose oxidase. The dissociation constant (KD) of the immobilized BM1 ligand with GAO was found to be equal to 45.8 μM, whereas the binding capacity was equal to 709 U per ml adsorbent. Therefore, the BM1 adsorbent was integrated in a facile two-step purification procedure for GAO. The purified enzyme showed a specific activity equal to 2038 U/mg, the highest reported so far, ∼74% overall recovery and a single band after sodium dodecylsulfate-polyacrylamide gel electrophoresis analysis. © 2002 Elsevier Science B.V. All rights reserved.	en
heal.journalName	Journal of Chromatography A	en
dc.identifier.issue	1-2	en
dc.identifier.volume	954	en
dc.identifier.doi	10.1016/S0021-9673(02)00176-0	en
dc.identifier.spage	137	en
dc.identifier.epage	150	en