Arabitol is a sugar alcohol, stereoisomer to xylitol, which is enlisted among the main target for biorefineries. It can serve as low calorie sweetener and as building block in the enantiopure synthesis of immunosuppressive glycolipids, herbicides, and drugs. Several studies described the fermentative production of arabitol by osmophilic yeasts, cultured with high concentrations of D-glucose. The utilization of cheaper carbon sources, such as glycerol or lactose, is of great interest for biorefinery implementation, but information on exploitation to arabitol production is still scarce. In the present study 50 yeasts belonging to 24 ascomycetous species were screened for the ability to grow and produce arabitol in presence of 80 g/L lactose or glycerol. Production from lactose was generally unsuccessful, the best producer being Kluyveromyces lactis WC 1401 with 0.94 g/L in 160 h. Production from glycerol was promising, with Zygosaccharomyces rouxii WC 1206, Pichia guilliermondii CBS 566, Hansenula anomala WC 1501, and Candida freyschussii ATCC 18737 yielding 3 to 4.5 g/L arabitol, with conversion yield (YP/S) ranging from 11 to 21.7%. Batch growth with high initial glycerol amount (160 g/L) resulted in higher production, with H. anomala WC 1501 yielding 10.0 g/L arabitol (YP/S = 12%) in 160 h. Preliminary bioreactor fermentations with H. anomala WC 1501 indicated that production is not growth associated and revealed some major parameters affecting production, such as the pH and the C:N ratio, that will be the target of following studies aiming at process optimization. Cultivation under controlled oxygenation (DOT = 20%) and pH (= 3.0) resulted in improvement in the performance of H. anomala WC 1501, yielding16.1 g/L arabitol. Cultivation in a medium with high C:N ratio, lacking inorganic nitrogen yielded 17.1 g/L arabitol. Therefore, this strain was selected for the development of a fed-batch process, aiming to improve the efficiency of the biomass, generated in the growth phase, and increasing the production in the stationary phase.