The study focus was an examination of the hydrothermal pretreatment method applied to the lignocellulosic substrate, represented by the prairie cord grass, and comparison between different conditions based on the yield of glucose after enzymatic hydrolysis. The treatment did not involve any chemicals usage. Enzymatic hydrolysis was performed in order to examine the amount of glucose which was released from pretreated materials. The most efficient pretreatment conditions were at high temperature and relatively short reaction time (210°C and 10 min), after which the lignocellulose structure was the most available for enzymes actions which resulted in a pretreatment conversion rate of 97%. Temperature had a significant influence on glucose release during the hydrolysis, which was confirmed by the Michaelis-Menten and kinetic models. Kinetic models were used to fit the inhibitors and their conversion rates were related to temperature.