AIM: Production of biofuel from lignocellulosic biomass has been extensively investigated in the world. The main issue in converting lignocellulosic biomass to fuel ethanol is the accessibility of the polysaccharides for enzymatic breakdown into monosaccharides. The aim of this article was evaluation of the physicochemical properties of steam exploded canola straw and its efficiency for enhancing fermentable sugar production. METHODS: In this study the effect of steam explosion on the physicochemical Characterization of Canola Straw was investigated. RESULTS: Steam exploded canola straw had higher Ash, acid detergent fiber (ADF), neutral detergent fiber (NDF) and cellulose than untreated canola straw while lower in hemicellulose in comparison with blank samples. By application of steam explosion treatment cellulose accessibility and it hydrolysis by enzyme hydrolysis increased. Maximum cellulose to glucose conversion (29.37%) was obtained for steam exploded sample while control samples showed 11.59% glucose yields. CONCLUSION: The effectiveness of steam explosion pretreatment in biomass conversion to fermentable sugar is due to its ability to change the biomass to sponge like and pores structure with higher porosity and lower bulk density in comparison with untreated canola straw that increased cellulose accessibility and it hydrolysis by enzyme hydrolysis.