Although organosolv pretreatment removed substantial amounts of lignin and xylan, the yield of glucan which is a major sugar source for fermentation to ethanol is more than 90% in most conditions of the organosolv pretreatment. Relative lignin contents of all pretreated biomass were more than 200 g kg<SUP>-1</SUP>, however enzymatic conversions were increased dramatically comparing to untreated biomass. Therefore the correlation between lignin and enzymatic hydrolysis could not be explained just by lignin content, and other changes resulting from lignin removal affected enzymatic hydrolysis. Results on enzymatic conversion and sugar recovery suggested that the critical temperature improving enzymatic hydrolysis significantly was between 120 <SUP>o</SUP>C and 130 <SUP>o</SUP>C. Microscopic analysis using Field emission scanning electron microscopy (FE-SEM) showed that structural lignin changes happened through organosolv pretreatment. Lignins were isolated from lignin carbohydrate complex (LCC) at the initial stage and then migrated to the surface of biomass. The isolated and migrated lignins were finally redistributed onto surface. These structural changes formed droplets on surface and increased pore volume in pretreated biomass. The increase in pore volume also increased available surface area and enzyme adsorption at initial stage, and thus enzymatic conversion increased significantly through organosolv pretreatment. It was verified that the droplets were mainly composed of lignin and the lignin droplets inhibited enzymatic hydrolysis through adsorption with cellulase.