This study explores utilizing pineapple peel (PP) hydrolysate as a promising carbon source for xylitol production, covering scopes from the pre-treatment to the fermentation process. The highest xylose concentration achieved was around 20 g/L via mild acid hydrolysis (5% nitric acid, 105 °C, 20-min residence time) with a solid loading of 10%. Two sets fermentability experiments were carried out of varying pH levels in synthetic media that includes acetic acid as the main inhibitors and hydrolysate supplemented with diverse nitrogen source. The results revealed that pH 7 exhibited the highest xylitol production, yielding 0.35 g/g. Furthermore, urea was found to be a highly promising and cost-effective substitute for yeast extract, as it yielded a comparable xylitol production of 0.31 g/g with marginal difference of only 0.01 g/g compared to yeast extract further highlights the viability of urea as the preferred option for reducing xylitol production cost. The absence of a significant difference between the synthetic media and hydrolysate, with only a marginal variance of 0.35 to 0.32 g/g, implies that acetic acid is indeed the primary constraint in xylitol production using PP hydrolysate. The study sheds light on PP biomass's potential for xylitol production, aligning economic benefits with environmental sustainability and waste management.