ABSTRACT
A rapid and efficient enzymatic procedure for the preparation of an immobilized ß-galactosidase has been described. In a first step, soybean peroxidase was used to catalyze the polymerization of a strategically activated phenol (N-Succinimidyl 3-(4-hydroxyphenyl)propionate, known as Bolton-Hunter reagent). The phenolic support was directly employed for immobilizing ß-galactosidase from Bacillus circulans (ATCC 31382, ß-Gal-3), giving rise to a new biocatalyst subsequently applied in the synthesis of a ß-galatodisaccharide (Gal-ß(1-3)-GlcNAc and Gal-ß(1-3)-GalNAc). The reaction proceeded with high conversion rates and total regioselectivity. Reusability assays were performed with the same reaction conditions finding that the immobilized enzyme retains about 55% of its activity after eight batches. Finally and based on our results, the two-step enzymatic procedure presented here is a good and green alternative to the preparation of carbohydrates with biological activities.