title: Affinity chromatography of beta-glucosidase and endo-beta-glucanase from Aspergillus niger on concanavalin A-Sepharose: implications for cellulase component purification and immobilization.
abstract: Affinity chromatography of a commercial preparation of beta-glucosidase from Aspergillus niger using concanavalin A-Sepharose (CAS) was employed as a means of purifying this glycoprotein. However, mannose (up to 1.08 M) was ineffective as an eluent of this enzyme from CAS, as were several other sugars and their derivatives, including 0.5 M glucose. Also, washing the CAS: beta-glucosidase complex with buffer at pH 3.5 in the absence of MnCl2 and CaCl2 (required to preserve the binding activity of concanavalin A below pH 5.0) did not result in elution of this enzyme. On the contrary, endoglucanase activity present in a crude cellulase complex (A. niger) which bound to CAS could be eluted by mannose (0.5-0.7 M) and was fractionated into at least two components. The CAS: beta-glucosidase complex hydrolyzed cellobiose to glucose and possessed an activity of 2,158 units/g dry CAS. It could be used, therefore, for continuous cellobiose hydrolysis without leakage of enzyme from the support.
mesh_terms: D000327:Adsorption; D001234:Aspergillus niger / Q000201:enzymology*; D002122:Calcium Chloride / Q000494:pharmacology; D002480:Cellulase / Q000302:isolation & purification*; D002712:Chlorides; D002846:Chromatography, Affinity; D003208:Concanavalin A; D004800:Enzymes, Immobilized; D005959:Glucosidases / Q000302:isolation & purification*; D008345:Manganese / Q000494:pharmacology; D017895:Manganese Compounds; D001617:beta-Glucosidase / Q000302:isolation & purification*
chemical_list: D002712:Chlorides; D004800:Enzymes, Immobilized; D017895:Manganese Compounds; D003208:Concanavalin A; D008345:Manganese; D005959:Glucosidases; D001617:beta-Glucosidase; D002480:Cellulase; D002122:Calcium Chloride; C025340:manganese chloride