The effects of several surfactants on the biosynthesis of beta-1,3-D-glucan (beta-glucan) and pullulan by Aureobasidium pullulans CCTCC M 2012259 were investigated, and Triton X-100 was found to decrease biomass formation but increase beta-glucan and pullulan production. The addition of 5 g/L Triton X-100 to the fermentation medium and bioconversion broth significantly increased beta-glucan production by 76.6% and 69.9%, respectively, when compared to the control without surfactant addition. To reveal the physiological mechanism underlying the effect of Triton X-100 on polysaccharides production, the cell morphology and viability, membrane permeability, key enzyme activities, and intracellular levels of UDPG, NADH, and ATP were determined. The results indicated that Triton X-100 increased the activities of key enzymes involved in beta-glucan and pullulan biosynthesis, improved intracellular UDPG and energy supply, and accelerated the transportation rate of precursors across the cell membrane, all of which contributed to the enhanced production of beta-glucan and pullulan. Moreover, a two-stage culture strategy with combined processes of batch fermentation and bioconversion was applied, and co-production of beta-glucan and pullulan in the presence of 5 g/L Triton X-100 additions was further improved. The present study not only provides insights into the effect of surfactant on beta-glucan and pullulan production but also presents a feasible approach for efficient production of analogue exopolysaccharides. Key points Triton X-100 increased beta-glucan and pullulan production under either batch fermentation or bioconversion. Triton X-100 increased the permeability of cell membrane and accelerated the transportation rate of precursors across cell membrane. Activities of key enzymes involved in beta-glucan and pullulan biosynthesis were increased in the presence of Triton X-100. Intracellular UDPG levels and energy supply were improved by Triton X-100 addition.