Considerable progress has been achieved for the production of bioethanol from lignocellulosic biomass. However, increasing the substrate concentration has been shown to decrease ethanol productivity. Therefore, Saccharomyces cerevisiae (S. cerevisiae) and Pichia sp. were used for ethanol production from glucose and xylose sugars in the present investigation. Furthermore, coculture fermentations were conducted for 13 g/L of sugar (10 g of glucose and 3 g/L of xylose). It was further used for ethanol production from ultrasonicationassisted NaOH (UA-NaOH) pretreated and enzymatically saccharified biomass in batch and fed-batch fermentation conditions. Furthermore, fed-batch fermentation was used for separate hydrolysis and cofermentation (SHCF) and simultaneous saccharification and cofermentation (SSCF) in shake flask conditions. The highest ethanol production of 12.2 and 7.9 g/L was observed for fedbatch SSCF deenanath grass (DG) and hybrid Napier grass (HNG) (Palkonal MBW as the enzyme) biomass (80 g/L), respectively, in shake flask conditions. However, increasing the biomass concentration to 270 g/L produced an ethanol concentration of 77.6 and 51.3 g/L for DG and HNG, respectively, in fedbatch SSCF conditions in a bioreactor. Furthermore, nuclear magnetic resonance studies of the residual biomass of DG revealed lower carbohydrate content, demonstrating the efficiency of the fermentation strategy. Life cycle analysis of DG and HNG revealed DG had a higher yield as well as lower List of Abbreviations: %, percent; 0 C, degree centigrade; 13 C, an isotope of carbon of mass number 13;