Sucrose phosphorylase (SP) is a useful biocatalyst for the selective transfer of alpha-glucosyl residues to a variety of acceptor molecules. Its industrial application is, however, hampered by the lack of enzyme variants that can withstand the process temperature of 60 degrees C. We have recently shown that the stability of the SP from Bifidobacterium adolescentis can be improved by immobilization on Sepabeads EC-HFA, and have now applied this biocatalyst for the continuous production of alpha-D-glucose-1-phosphate from sucrose. To lower the costs, the enzyme has only been partially purified prior to immobilization. Interestingly, the presence of substrate was found to dramatically enhance the stability of the biocatalyst, allowing its use in a packed-bed reactor for more than 2 weeks at 60 degrees C without loss of activity. The overall process generated a space-time yield of 179 g/l/h, and the product could be recovered in crystalline form with a yield of 86%. (C) 2011 Elsevier Ltd. All rights reserved.