Hemicellulosic hydrolyzates from vineshoot trimmings obtained by dilute sulfuric acid hydrolysis were evaluated for xylitol production by Debaryomyces hansenii NRRL Y-7426. Bioconversion was not efficient, however, since a mixture of products (mainly ethanol) was achieved. Taking into account that hexoses (such as glucose or mannose) can inhibit xylose metabolism by repression and inactivation of the xylose transport system or catabolic enzymes and that these hemicellulosic hydrolyzates are characterized by a high glucose concentration, a novel technology was developed, sequentially transforming glucose into lactic acid by Lactobacillus rhamnosus followed by fermentation of xylose into xylitol by Debaryomyces hansenii after L. rhamnosus removal by microfiltration. Optimal conditions were achieved using detoxified concentrated hemicellulosic hydrolyzates, after CaCO 3 addition in both stages of fermentation and using nitrogen purges after sampling in order to reduce the oxygen dissolved. Under these conditions 31.