Declining supplies of fossil fuels, increasing population, global industrialization and demand for transportation fuels has triggered an increase in the demand for renewable energy sources. To address such problems most of the green research in the recent years has focused on the development of bioethanol (23 MJ/L) as a substitute to conventional gasoline (34.3 MJ/L) based fuels owing to the similarity in energy density values in addition to several other advantages (American Council on renewable energy, 2010). Second-generation biofuels are derived from lignocellulosic biomass or woody crops, mostly coming from agricultural residues. Extraction of fuel from such biomass is difficult because of their recalcitrant nature (corn stover, rice straw, wheat straw, sugar cane and sweet sorghum). Lignocellulosic fuel has the potential to solve several problems (food competing with fuel) that are currently associated with first generation biofuels. Moreover, lignocellulosic fuels can supply a larger proportion of the global fuel leading to sustainability at lower cost, and with greater environmental benefits (Liz Marsall, 2009). The production of ethanol from the complex sugars in leaves and stalks is a promising strategy to radically broaden the range of possible ethanol feedstock. Keywords: lignocellulose, bioethanol, biomass, pretreatment, hydrolysis, fermentation.

Lignocellulosic biomass is one of the largest carbohydrate sources and has huge potential for biofuels production. However, the problem with lignocellulosic feedstock is that it has useful sugars locked in by lignin, hemicellulose, and cellulose. Some kind of pretreatment; therefore is needed to make carbohydrate accessible which later can be fermented to produce ethanol. The results from this research indicated that the yields of glucan (93%) and xylan (82.8%) were improved by using milling combined with ELLA pretreatment. The optimal enzymatic hydrolysis efficiencies were obtained under 10 min for ball milling time, pretreatment at 1 h, temperature at 150°C, S/L = 0.5 and ammonia loading at 0.25 g-NH3/g-biomass. This method reduced the pretreatment time and short milling time and thus has potential of reducing the energy consumption and promising the application in the large scale. Keywords: corn stover, milling pretreatments, ammonia pretreatment, bioethanol.