Abstract. Pervaporation, a membrane-based technique, is taken into consideration in order to separate water from biodiesel-methanol mixtures. Several operational conditions in biodiesel production could cause water contamination into the reaction mixture, which can affect biodiesel production and quality. In this study, bacterial cellulose-alginate (BCA) nanocomposite film was applied as a selective membrane in order to separate water from biodiesel-methanol mixtures using pervaporation. For effective operation, factors that affected the performance in the pervaporation, such as concentration of water in the biodiesel-methanol mixtures and temperature of the process were investigated. It was demonstrated that, the BCA membrane has good potential for removing water from the biodiesel-methanol mixtures. Under a permeate pressure of 10 mmHg and 30 °C, the BCA membrane could separate water from the mixture containing methyl ester (C10:0): methanol: water at a weight ratio of 42.3: 52.7: 5 with a total permeate flux of 148 g/(m 2 h) and a water selectivity of 332. The permeate was contained 94.5% w/w water and methyl ester was completely rejected by the BCA membrane. The increase of water concentration in biodiesel-methanol mixtures and the temperature rise resulted in an increase in the permeate flux but lowered the selectivity.