The effect of sodium citrate on ε-PL fermentation of S. albulus was investigated. The results showed that sodium citrate concentrations had slight effect on cell growth, but significantly affect the synthesis of ε-PL. The addition of 2 g/L sodium citrate at 0 h resulted in the highest ε-PL concentration of 0.92 g/L. By adding 2 g/L sodium citrate at 0 h and 300 μg/L biotin at 36 h to fermentation media, the cell dry weight and ε-PL yield reached the highest values of 7.86 g/L and 1.10 g/L, respectively, being 1.30 and 1.93 folds to the control respectively. It was demonstrated that sodium citrate and biotin will promote cell growth and ε-PL yield of fermentation of S. albulus.