Cold enzymatic hydrolysis conditions for bioethanol production were optimized using multi-objective optimization. Response surface methodology was used to optimize the effects of α-amylase, glucoamylase, liquefaction temperature and liquefaction time on S. cerevisiae biomass, ethanol concentration and starch utilization ratio. The optimum hydrolysis conditions were: 224IU/g starch α-amylase, 694IU/g starch glucoamylase, 77°C and 104min for biomass; 264IU/g starch α-amylase, 392IU/g starch glucoamylase, 60°C and 85min for ethanol concentration; 214IU/g starch α-amylase, 398IU/g starch glucoamylase, 79°C and 117min for starch utilization ratio. The hydrolysis conditions were subsequently evaluated by multi-objectives optimization utilizing the weighted coefficient methods. The Pareto solutions for biomass (3.655–4.380×10 8 cells/ml), ethanol concentration (15.96–18.25wt.%) and starch utilization ratio (92.50–94.64%) were obtained. The optimized conditions were shown to be feasible and reliable through verification tests. This kind of multi-objective optimization is of potential importance in industrial bioethanol production.