Efficient utilization of lignocellulosic feedstocks (agricultural wastes) offers an opportunity to reduce the cost of producing fuel ethanol. However, no organism has been found in nature which can rapidly and efficiently metabolize different sugars from hydrolyzed lignocellulose into ethanol. Recombinant Escherichia coli strain rC13 fermenting C5 and C6 sugars to ethanol was constructed. The effect of some factors such as minerals, pH, and initial biomass on the ethanol producibility of recombinant E. coli strain was investigated. Ethanol producibilities of rC13 strain were significant higher in 0.2 M buffering postassium phosphate (pH 7.0) minimum medium (M9) adding 0.05 percent minerals (MgCl2, CaCl2, FeSO4.7H2O). The ethanol producibility of rC13 strain using different sugars such as C5 (xylose, arabinose) and C6 (glucose, mannose) was also investigated. Ethanol concentrations in fermented medium from C5 sugars were lower than that from C6 sugars (3.16 percent and 3.94 percent, respectively). .Recombinant strain was able to ferment hydrolysate of sugarcane bagasse.