title: Affinity chromatography of invertase on concanavalin A-bead cellulose matrix: the case of an extraordinary strong binding glycoenzyme.
abstract: This work presents confirmation of the biospecific character of the interaction of Concanavalin A (Con A) immobilized on bead cellulose with invertase. In spite of the extraordinary strong binding of invertase to this Con A conjugate (KD = 5 x 10(-9) M), conditions have been found to use Con A-bead cellulose as an affinity chromatography medium. The effective factor in the release of the bound invertase by the counter ligand (alpha-methyl-D-mannopyranoside) is the time of incubation. This phenomenon was demonstrated in both batch and flow-through experiments. A concentration of 1.5 mg Con A per ml of gel was found to be suitable with regard to the maximal invertase/Con A binding ratio and the optimal invertase recovery (94%). As a result of the strong biospecific interaction the purification of invertase was very effective (above ten-fold). Verification by FPLC and PAGE of the product purity revealed only one significant protein band.
authors.affiliation: Institute of Chemistry, Slovak Academy of Sciences, Bratislava.
authors.forename: D
authors.initials: D
authors.lastname: Mislovicová
authors.forename: M
authors.initials: M
authors.lastname: Chudinová
authors.forename: P
authors.initials: P
authors.lastname: Gemeiner
authors.forename: P
authors.initials: P
authors.lastname: Docolomanský
mesh_terms: D002482:Cellulose / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000379:methods*; D003208:Concanavalin A / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D011485:Protein Binding; D043324:beta-Fructofuranosidase
chemical_list: D003208:Concanavalin A; D002482:Cellulose; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase