Ozonated energy grass varieties were enzymatically hydrolyzed to establish process parameters for maximum fermentable sugar production. Conditions for ozonolysis were selected on the basis of maximum delignification and glucan retention after pretreatment. To study the effect of lignin degradation products generated during ozonolysis on cellulolytic enzymes, hydrolysis was carried out for washed and unwashed pretreated solids. Washing the solids significantly (p<0.05) enhanced glucan conversion from 34.3% to 100% while delivering glucose yields of 146.2-431.9 mg/g biomass. Highest fermentable sugars were produced when grasses were ozonated for maximum delignification and washed solids were hydrolyzed using 0.1 g/g Cellic (R) CTec2. In a comparative study on alkaline pretreatment with 1% NaOH for 60 min, Saccharum arundinaceum exhibited the highest glucan conversion with maximum sugar production of 467.9 mg/g. Although ozonolysis is an effective and environmentally friendly technique for cellulosic sugar production, process optimization is needed to ascertain economic feasibility of the process. (C) 2013 Elsevier Ltd. All rights reserved.