Production of Bioethanol from lignocellulosic biomass is a potential source of energy to fulfill current demands as well as in reducing green house gas emission. The microbial preparation of cellulase / xylanase can stand as an achievement that addresses the alternative energy need of today’s world, especially in landlocked countries like ours, where we are totally dependent to other countries for fuels which could lead new industrial revolution. This process of bioethanol production generally involves hydrolysis of lignocellulosic biomass to fermentable sugars, and fermentation of the sugars to ethanol. Different microflora from cellulolytic microbes in the soil sample were isolated and sub-cultured, on the basis of their ability to secret a wide variety of plant hydrolyzing enzymes. The samples were given name as Lc, SNK, P1D, P2D, PSS, PCR, SS, respectively. The congo red test on the samples also showed greatest zone of inhibition in SNK, Lc, P1D and PSS thus implying greater hydrolytic capacity. The broth culture of the isolated and identified organisms were used for the extraction of extracellular enzymes via centrifugation at 2000 rpm for 20 minutes. Enzymatic assay like CMCase, Xylanase and β-glucosidases assay showed the highest activity of enzymes extracted from sample PSS, P1D and SNK. Studies of optical density at 540 nm after DNS reagent test resulted the finding that PSS has both  greater cellulolytic and Xylanolytic activity comparatively. The optimized pretreatment strategy (2% NaOH with 1:50 substrate ratio) was used to make cellulose and xylan accessible for enzymatic digestion. Saccharification was carried out by adding crude enzymes extracted from samples to pretreated substrate sugarcane bagasse. After saccharification aliquots from each tube was taken for redcuing sugar estimation by DNS method. The samples sent to macrogen korea for 16s rRNA full sequencing suggested that samples SNK, PSS and Lc were Bacillus sps, Staphylococcus sps and Bacillus sps respectively by BlastN.

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