Bioethanol is in limelight these days being safe and renewable. Its production from plant feed stock is getting considerable importance as it does not pose food competition. Therefore, current study was focused on the enzymatic conversion of wheat straw in to fermentable saccharides which can further be used for bioethanol formation. Thermophilic cellulases i.e. Endo-1,4-ß-glucanase, Exo-1,4-ß-glucanase and ß-1,4-glucosidase were used for this purpose. Differently pretreated i.e. acid and alkali wheat straw samples were analysed for their effective enzymatic conversion to less complex sugars. Two different strategies (Simultaneous and Sequential) for cellulase action on substrate were assessed. Furthermore, effect of different reaction conditions such as buffer, pH, temperature and substrate concentration was also evaluated. Maximum saccharification i.e. 43.78% (p<0.05) have been recorded using alkali treated wheat straw substrate in a concentration of 1.5% (w/v) by sequential addition of cellulases in a citrate phosphate buffer of pH 7 at an incubation temperature of 55°C.The microscopic examination of saccharified biomass showed clear disruption in the cellulose fibres. Moreover, thin layer chromatography of saccharified slurry showed the presence of variable fermentable sachharides.