Hybrid affinity chromatography of alpha-galactosidase from Verbascum thapsus L.
Purification of alpha-galactosidase from the roots of Verbascum thapsus L. was difficult to achieve using conventional methods due to the presence of coloured contaminants. A newly developed procedure, hybrid affinity chromatography, which was based on a mixed matrix separation procedure, using a substrate analogue and an immobilized metal affinity matrix as ligands, respectively, allowed the purification of this enzyme with good recovery. The method should be applicable to other proteins as well.
Bom
I
I
Unilever Research Laboratory, Vlaardingen, The Netherlands.
van Wassenaar
D
D
Boot
J
J
D002846:Chromatography, Affinity / Q000379:methods*; D003208:Concanavalin A; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D037121:Plant Lectins; D018517:Plant Roots / Q000201:enzymology; D010944:Plants / Q000201:enzymology*; D000519:alpha-Galactosidase / Q000302:isolation & purification*
D037121:Plant Lectins; D003208:Concanavalin A; D000519:alpha-Galactosidase