The oil palm empty fruit bunches (EFB) are an attractive source of carbon for the production of biochemical products, therefore, the aim of this work is to analyze the effect of the steam explosion (SE) pretreatment under autocatalytic conditions on EFB using a full experimental design. Temperature and reaction time were the operational variables studied. The EFB treated at 195 °C for 6 min showed an increase of 34.69% in glycan (mostly cellulose), and a reduction of 68.12% in hemicelluloses, with increased enzymatic digestibility to 33% producing 4.2 g L−1 of glucose. Scanning electron micrographs of the steam treated EFB exhibited surface erosion and an increased fiber porosity. Fourier transform infrared spectroscopy showed the solubilization of hemicellulose and modification of cellulose in treated EFB.