In this study, endoglucanase was produced from Trichoderma harzianum in submerged fermentation using sugarcane as a substrate. For enhanced enzyme production, various parameters were optimized by one factor at a time approach. Maximum enzyme yield was obtained with initial medium pH of 4.0, inoculum size of 2%, sugarcane bagasse concentration of 2% and incubation temperature of 30ºC for 72 h of fermentation period. Addition of tween-80 (0.2%) to the fermentation medium had significant impact on endoglucanase production. Further supplementation of glucose and NaNO3 as synthetic carbon and nitrogen source favoured enzyme pro- duction respectively. Results of this study are very helpful in large scale enzyme production.