The aim of the present work was to use a thermophilic consortium for H-2 production using lignocellulosic biomass in a single pot. The thermophilic consortium, growing at 60 degrees C utilized both glucose and xylose, making it an ideal source of microbes capable of utilizing and fermenting both hexose and pentose sugars. The optimization of pH, temperature, and substrate concentration increased the H-2 production from 1.07 mmol H-2/g of prairie cordgrass (PCG) to 2.2 mmol H-2/g PCG by using the thermophilic consortium. A sequential cultivation of a thermostable lignocellulolytic enzyme producing strain Geobacillus sp. strain WSUCF1 (aerobic) with the thermophilic consortium (anaerobic) further increased H-2 production with PCG 3-fold (3.74 mmol H-2/g PCG). A single pot sequential culturing of aerobic and anaerobic microbes can be sustainable and advantageous for industrial scale production of biofuels.