3.4. Release of α-lFucosidase

Cell pellets, obtained as described above, were washed twice with isotonic solution (NaCl 0.9%). The cells were added to a solution containing 0.5 g/L of pNP-Fuc and incubated at 37 °C for 48 h. Samples were taken periodically and centrifuged at 3110× g for 20 min at 4 °C. Both supernatant and cellular pellet were resuspended in phosphate buffer (100 mM, pH 7) and analyzed for α-lfucosidase activity. Released enzyme in the supernatant was used for the synthesis of fucosyl-oligosaccharides.