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Wheat straw was pretreated by combined mechanical destruction and alkaline pretreatments to enhance enzymatic saccharification. Four strategies were employed to evaluate the potential of wheat straw as a feedstock for fermentable sugar production. The effects of the pretreatments on the substrate morphology, size distribution, chemical composition, and cellulose crystallinity, along with the subsequent enzymatic digestibility, were investigated. Optical microscope images showed that mechanical pretreatment alone resulted in poor fiber defibrillation, wherein samples mostly consisted of rigid fiber bundles, while integrated mechanical destruction and alkaline pretreatment led to relatively good fiber defibrillation. Low temperature NaOH/urea pretreatment can fibrillate the rigid fiber bundles into a relatively loose network and alter the structure of the treated substrate to make cellulose more accessible. The glucan conversion rates were 77% and 95% for integrated mechanical destruction and alkaline pretreatments and mechanical destruction followed by low temperature NaOH/urea and ammonium/urea pretreatments, respectively, after 72 h of enzymatic hydrolysis with enzyme loadings of 10 FPU cellulase per g of oven-dry substrate. [ABSTRACT FROM AUTHOR]