A rapid and efficient enzymatic procedure for the preparation of an immobilized β-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 β–galactosidase from Bacillus circulans (ATCC 31382, β–Gal–3), giving rise to a new biocatalyst subsequently applied in the synthesis of a β–galatodisaccharide (Gal–β(1–3)–GlcNAc and Gal–β(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. Fil: Levin, Gustavo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Entre Ríos. Universidad Nacional de Entre Ríos. Centro de Investigaciones y Transferencia de Entre Ríos; Argentina. Universidad Complutense de Madrid; España Fil: Gómez, Sara. Universidad Complutense de Madrid; España Fil: Glodowsky, Alejandro Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Entre Ríos. Universidad Nacional de Entre Ríos. Centro de Investigaciones y Transferencia de Entre Ríos; Argentina Fil: Cascone, Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina Fil: Hernáiz Gómez-dégano, María Jose. Universidad Complutense de Madrid; España