Bioethanol production using lignocellulosic materials (the second-generation technology) was consist of pretreatment and simultaneous saccharification and fermentation (SSF) process. This research optimize the SSF incubation time using cellulose enzyme and Saccharomyces cerevisiae combination for sugarcane bagasse. The bagasse was pre-treated by delignification with NaOH. The SSF was conducted by Sacharomyches cereviceae, cellulose enzymes (Trichoderma reesei) and nutrients combination during 3 d, 5 d and 7 d as incubation time variables. The optimum result was 0.748 5 % of ethanol concentration or dry weight conversion by 11.810 5 g · L–1 which obtained after 5 d incubation time based on ethanol conversion, microbiology test and sugar reduction concentration.