Sugarcane bagasse (SCB) is an abundant by-product from sugar production and promising biomass for cellulose extraction. Simulated elephant colon pretreatment (SEP) to reduce chemical use in cellulose extraction from SCB was investigated using elephant dung as fermentation inoculum. The 16S rRNA gene sequences showed microorganisms in elephant dung that corresponded to metabolites during pretreatment. Organic acid accumulation in the fermentation broth was confirmed by the presence of lactic, acetic, propionic and butyric acids. Lignin peroxidase, manganese peroxidase and xylanase detected during the pretreatment enhanced lignin removal. The SEP fiber showed increased cellulose content, while lignin content decreased with reduced bleaching time from 7 to 5 h and high whiteness and crystallinity indices. Lignin removal was also confirmed by Fourier transform infrared spectroscopy. Scanning electron microscopy revealed increasing internal surface area through opening up the fiber structure. SEP offered an efficient and promising approach for cellulose fiber extraction with reduced use of chemicals for the bleaching process.