Purification of glucoamylase by acarbose (BAY g-5421) affinity chromatography.
Aspergillus niger and Rhizopus sp. glucoamylases were purified on an affinity chromatography column from commercially available, impure enzyme preparations. Up to 2 mg of glucoamylase protein was bound without leakage to a 1-ml affinity gel column (0.7 X 2.5 cm) possessing a covalently linked acarbose ligand (1 mg acarbose/g wet gel), and the bound enzyme was specifically released by irrigation of the column with a solution of maltose. A complete cycle of purification was accomplished in about 8 h. Glucoamylases were recovered, in more than 80% yield, free of alpha-amylase activity and possessing specific activities comparable to those of preparations obtained by time-consuming, multistep procedures involving several ion-exchange and hydrophobic column fractionations. Thus, acarbose affinity chromatography provides a general method for the rapid and efficient purification of the glucoamylases, and seems to be ideally suited for scale-up for the commercial purification of these enzymes.
D020909:Acarbose; D000818:Animals; D001234:Aspergillus niger / Q000201:enzymology*; D002846:Chromatography, Affinity / Q000379:methods; D005087:Glucan 1,4-alpha-Glucosidase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D005959:Glucosidases / Q000302:isolation & purification*; D006026:Glycoside Hydrolases / Q000302:isolation & purification; D007700:Kinetics; D010179:Pancreas / Q000201:enzymology; D010944:Plants / Q000201:enzymology; D012233:Rhizopus / Q000201:enzymology*; D013295:Streptococcus mutans / Q000201:enzymology; D013552:Swine; D014312:Trisaccharides
D014312:Trisaccharides; D005959:Glucosidases; D006026:Glycoside Hydrolases; D005087:Glucan 1,4-alpha-Glucosidase; D020909:Acarbose