Characterization of the immobilized beta-galactosidase C from Bacillus circulans and the production of beta(1-3)-linked disaccharides

• Published in: Bioscience, biotechnology, and biochemistry Vol. 62; no. 7; pp. 1313 - 1317
• Main Authors: Naundorf, A. (Meiji Milk Products Co. Ltd., Odawara, Kanagawa (Japan). Inst. of Health Science), Caussette, M, Ajisaka, K
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 1998; Japan Society for Bioscience Biotechnology and Agrochemistry
• Subjects: BACILLUS; Bacillus circulans; BETA GALACTOSIDASA; BETA GALACTOSIDASE; Biological and medical sciences; Biotechnology; ENZIMAS INMOVILIZADAS; ENZYME IMMOBILISEE; Fundamental and applied biological sciences. Psychology; Gal-β(1→3)-GalNAc; immobilization; Immobilization of enzymes and other molecules; Immobilization techniques; IMMOBILIZED ENZYMES; Methods. Procedures. Technologies; OLIGOSACARIDOS; OLIGOSACCHARIDE; OLIGOSACCHARIDES; transglycosylation; β-galactosidase; Enzymatic synthesis; Bacillus circulans; Enzyme; Immobilization; Bacillaceae; Disaccharide; Thermal stability; Transglycosylation; Organic solvent; Bacillales; Glycosidases; Biotransformation; Production; Bacteria; Hydrolases; O-Glycosidases; Immobilized enzyme; β-Galactosidase; β-galactosidase; immobilization; transglycosylation; Gal-β(1→3)-GalNAc
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.62.1313

A recombinant beta-galactosidase, which was obtained from the beta-galactosidase C gene of Bacillus circulans and cleaves the non-reducing end galactosyl residue of beta(1 to 3)-linkages selectively, was immobilized using CNBr-Sepharose. Although the effect of pH was not changed by the immobilization, the thermostability and stability in the presence of DMF were increased. Optimization of the transglycosylation using para-nitrophenyl beta-d-galactopyranoside as a donor and benzyl-alpha-D-N-acetylgalactosaminide as an acceptor afforded a beta(1 to 3)-linked disaccharide derivative with 62% molar yield in a gram scale synthesis. Using the methyl-analogue as an acceptor, 53% of the acceptor was converted to the respective beta(1 to 3)-disaccharide