Yeast immobilization offers operational advantages such as high cell concentration, and some drawbacks related to cell leaking and restricted mass transfer inside particles. The influence of bead size, chitosan, bead charge, volume of liquid media, and the use of corncob hydrolyzates and vinasses as culture medium were analyzed on xylitol production by Debaryomyces hansenii immobilized in alginate beads. The results showed a profuse growth of free cells, accounting 75-95% of total biomass, but electron micrographs revealed the generation of a dense biofilm with hyphal morphology at the bead surface and a very low intraparticular growth. Xylitol production was not affected by the size of particle; however chitosan had a negative effect. The use of corn cob as carbon source and twofold diluted vinasses as economic nutrients incremented xylitol concentration to 13.7 g L-1 (Y-P/S = 0.56 g g(-1); Q(P) = 0.29 g L-1 h(-1)). The best conditions corresponded to high bead charges and intermediate liquid volumes (44 g Na-alginate and 110 mL liquid medium). These results showed the feasibility of employing these cheap substrates, reflected the importance of the microaerobical conditions, and pointed to the favorable effect of cell immobilization on the metabolism of xylitol production. (C) 2013 Elsevier Ltd. All rights reserved.