Biocellulose produced by Acetobacter xylinum displays unique properties far and better than the plant cellulose, including a unique nanostructure, high water holding capacity, degree of polymerisation, mechanical strength, and crystallisation. The production of biocellulose, however, is costly due to the carbon sources and fermentation feedstocks. Pineapple peel waste has potential to be used as the carbon source since it is cheap and easy to obtain. This study was conducted to determine the composition of reducing sugar in pineapple peel using the high performance liquid chromatograph (HPLC) to produce and compare biocellulose derived from pineapple peel with biocellulose derived from commercial Hestrin-Schramm (HS) medium and other agricultural wastes. Moreover, this study also aimed to produce the process flow diagram and process and instrumentation diagram for large production of biocellulose using pineapple peel. Pineapple peel has approximately 0.58 g/mL of glucose and 0.78 g/mL of fructose, which is suitable to be used as fermentation medium for biocellulose production by Acetobacter xylinum. The dry weight of biocellulose that was produced using pineapple peel was 15.67% heavier than biocellulose produced using HS medium. Based on findings of the current study, it can be concluded that pineapple peel is suitable to be used as fermentation medium for high production of biocellulose.