The optimal mixed culture model of oleaginous yeast Rhodotorula glutinis and microalga Chlorella vulgaris was confirmed to enhance lipid production. A double system bubble column photo-bioreactor was designed and used for demonstrating the relationship of yeast and alga in mixed culture. The results showed that using the log-phase cultures of yeast and alga as seeds for mixed culture, the improvements of biomass and lipid yields reached 17.3% and 70.9%, respectively, compared with those of monocultures. Growth curves of two species were confirmed in the double system bubble column photo-bioreactor, and the second growth of yeast was observed during 36-48 h of mixed culture. Synergistic effects of two species for cell growth and lipid accumulation were demonstrated on O-2/CO2 balance, substance exchange, dissolved oxygen and pH adjustment in mixed culture. This study provided a theoretical basis and culture model for producing lipids by mixed culture in place of monoculture. (C) 2014 Elsevier Ltd. All rights reserved.