Energy production and consumption is a major environmental and economic concern in the world today. Biofuel produced from lignocellulosic biomass is being explored as an alternative energy source to petroleum. Lignocellulosic biomass is comprised of cellulose, hemicellulose, and lignin. To produce biofuels, lignocellulosic biomass must be broken down to expose cellulose for further pretreatment. Pretreatment of lignocellulosic biomass is an essential step in the process because lignin is hypothesized to lower the efficiency of enzymatic hydrolysis. The focus of this study was to understand how different compositions of lignin affects efficiency of enzymatic hydrolysis. Lignin was extracted from Oregon White Oak (Quercus garryana), Western Red Cedar (Thuja plicata), and Tall Fescue (Festuca arundinacea) using an organosolv pretreatment. Then a dilute acid pretreatment was used to impregnated the different types of lignin onto three types of pure cellulose: Avicel, bacterial cellulose and carboxymethyl cellulose (Simonsen Lab, Oregon State University). Enzymatic hydrolysis was performed in a small-scale 96-well plate at 59°C for 72 hours. Scanning Electron Microscopy (SEM) was used to analyze the lignin droplets at all stages of pretreatment and hydrolysis. As expected the lignin from Oregon White Oak (Hardwood) and Tall Fescue (Grass) behaved similarly in enzymatic hydrolysis efficiency, structural appearance, and droplets size. Lignin from Western Red Cedar (Softwood) behaved differently compared to Oregon White Oak and Tall Fescue lignins as expected due to its different lignin composition and syringal/guaiacyl (S/G) ratio.