Production of an alternative sweetener, xylitol from oil palm empty fruit bunch (OPEFB) waste fiber (a palm oil mill waste) was studied in laboratory following combination of chemical and biotechnological processes. Xylose sugar was produced from oil palm empty fruit bunch fiber using sulfuric acid hydrolysis process under various operating conditions. The results show that production of xylose can be correlated with operating variables e.g. acid concentration, operating temperature and reaction time. On the other hand production of xylitol depends on concentration of xylose & presence of inhibitors in the acid hydrolysate; type of yeast culture and fermentation time period. This results indicated that high production of xylitol can be achieved within short period of fermentation by proper control of formation of inhibitors in the resulting acid hydrolysate and using C.guillermondii as fermentative yeast.