Xylitol is a sugar alcohol that has a commercial application as a sugar substitute. It can be produced by a number of species of the yeast Candida when grown on xylose as a carbon source. The objective of this study was to determine whether Candida tropicalis ATCC 750, Candida tropicalis ATCC 20216, Candida mogii ATCC 18364, Candida guilliermondii ATCC 20216 or Candida guilliermondii ATCC 201935 could utilize the xylose released following alkaline hydrogen peroxide-hydrolysis of the prairie grass Big Bluestem (Andropogon gerardii) to produce xylitol. The dried grass was ground, treated with an alkaline hydrogen peroxide solution for 24 hours and hydrolyzed with a thermostable xylanase.  The yeast cells were grown in a phosphate-buffered medium (pH 5.0) containing bluestem hydrolysate in shake flask cultures.  The strains were grown for 168 hours at 30^oC. A culture grown for 48 hours containing the same medium was used to inoculate each shake flask culture. The concentration of xylitol produced was measured spectrophotometrically using the enzyme sorbitol dehydrogenase. All the strains screened produced xylitol after growth on the hydrolysate medium. Of the strains tested, it was determined that C. mogii ATCC 18364 synthesized the highest xylitol level after 168 hours of growth on the medium. In contrast, the C. tropicalis strains produced the lowest xylitol concentration in the medium after 168 hours. In conclusion, it was noted that species of the yeast Candida could utilize hydrolyzed bluestem grass to support xylitol synthesis but the concentration of xylitol synthesized varied according to the species investigated.