Using high gravity fermentation for bioethanol production by Saccharomyces cerevisiae is faced with many challenges such as increasing osmotic stress and viscosity. Unfortunately, increasing specific gravity was accompanied by a decrease in fermentation efficiency. A two-stage sequential optimization strategy was carried out to solve this problem. The first stage was carried out using the Plackett-Burman Design in which eight factors were investigated at three molasses specific gravities (1.130, 1.145, 1.160). Urea, wheat bran, soy flour, and inoculum size were significant model terms and had positive effect on bioethanol production. When a validation test was performed using the predicted conditions, about a more than two-fold increase in fermentation efficiency (FE) was achieved (83%) at specific gravity 1.130, compared to the basal condition. The second stage (Taguchi Design) was conducted using eight factors. Urea, wheat bran, temperature, and agitation speed showed a significant effect on fermentation efficiency. A validation test was carried out and up to 83.82 % fermentation efficiency was recorded, which was very close to that achieved under normal gravity. In addition to the high value of FE, this strategy is cost-effective and time-saving.