Abstract Several strategies based on a two steps organosolv pretreatment followed by enzymatic hydrolysis of sugarcane bagasse (SCB) were evaluated with the objective of selecting operational conditions suitable to promote an efficient and low cost production of ethanol. Initially, the influence of six variables used for the organosolv pretreatment was studied. The variables included the time of the first organosolv pretreatment step, the use of 45% ethanol as pulping solution, solid-to-liquid ratio of the ethanol solution used during the first pretreatment step, time of second organosolv pretreatment, concentration of ethanol and concentration of NaOH solution used in the second pretreatment step. Further assays of enzymatic hydrolysis were carried out to promote additional reduction in the costs of the process and improve the results of cellulose conversion to glucose. Eliminating the milling step of the pretreated SCB, using a commercial tensoactive (composed of esters and several surfactants), and recycling 50% of the slurry obtained during the second step of organosolv pretreatment as reaction medium proved to be feasible for use during the enzymatic hydrolysis. Fermentation of the glucose medium produced under the selected pretreatment conditions to ethanol by Saccharomyces cerevisiae occurred with 81% efficiency and a cost of 102.88 $/hL of ethanol.