title: Separation of enzymes from polyenzyme mixture used in medicine and pharmacy. II. Purification and characterization of extracellular beta-glycosidases with high transglycosylation activities from Aspergillus oryzae.
abstract: Three extracellular beta-glycosidases with different substrate specificities have been isolated from Aspergillus oryzae (Luizym) and purified to electrophoretic homogeneity by molecular-sieve and ion-exchange chromatographic methods. The enzymes were characterized as monomeric glycoproteins with an estimated molecular mass of 95 kDa by SDS-PAGE and 92 kDa by gel-permeation chromatography on Superose 12 HR 10/30. beta-glycosidase I (pHopt 4.8; Topt 40 degrees C, pl 4.5) was able to catalyze the hydrolysis of aryl-beta-galactopyranosides (o- and p-), where as gamma-glycosidase II and III were found to be active towards aryl-beta-gluco- and xylopyranosides. The specific chemical modifications of different amino acid residues showed that tryptophyl and carboxyl residues play an important role for the enzyme activity. The isolated beta-glycosidases exhibited high levels of transglycosylation activities and were used for the production of tri- and tetrasaccharides from lactose and whey permeate.
authors.affiliation: Department of Biochemistry, Faculty of Biology, University of Sofia St. Kliment Ohridski, Bulgaria.
authors.forename: S D
authors.initials: SD
authors.lastname: Petrova
authors.forename: N G
authors.initials: NG
authors.lastname: Bakalova
authors.forename: E D
authors.initials: ED
authors.lastname: Bankova
authors.forename: S B
authors.initials: SB
authors.lastname: Andonova
authors.forename: D N
authors.initials: DN
authors.lastname: Kolev
mesh_terms: D001236:Aspergillus oryzae / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004798:Enzymes / Q000302:isolation & purification*; D006026:Glycoside Hydrolases / Q000302:isolation & purification*; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D007525:Isoelectric Focusing; D007700:Kinetics; D008670:Metals / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D004798:Enzymes; D007202:Indicators and Reagents; D008670:Metals; D006026:Glycoside Hydrolases