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
Petrova, S.D.; Bakalova, N.G.; Bankova, E.D.; Andonova, S.B.; Kolev, D.N.
Die Pharmazie 58(1): 63-67
2003
ISSN/ISBN: 0031-7144 PMID: 12622257 Document Number: 555516
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.