In order to reduce the cost of lignocellulosic ethanol production, a model was previously developed to predict the amount of fermentable sugar released based on parameters concerning both biomass and enzymes. To validate this model, enzymatic hydrolysis must be carried out using individual purified cellulase enzymes. Endo-1,4-β-D-glucanase I (EG I), a typically difficult enzyme to purify, was partially purified using three steps of fast protein liquid chromatography. The purity and identity were determined using SDS-PAGE and specific activity assays. The partially purified fractions of EG I, and three other purified cellulase enzymes were used to hydrolyze three different cellulose types. The three enzyme mixtures, with increasing fractions of EG I, showed insignificant differences for all three types of cellulose used.
Keywords: biofuels, lignocellulosic ethanol, protein purification, cellulase, enzymatic hydrolysis