Abstract: The influence factors ofβ- carotene production by co- fermentation of Rhodotorula glutinis and Rhodotorula RY- 01 were studied. The results showed that the production was significantly improved by co-fermentation of two strains than single strain fermentation. β- carotene production reached 8837.83μg /L and the yeast biomass was 23.56g /L.The accumulation of β- carotene was not synchronized with biomass accumulation, which was lag behind the biomass.The optimum carbon source and nitrogen source of co- fermentation medium was maltose and peptone respectively.The β-carotene production was not significantly improved by K+and Mg2 +.The extract of orange peel powder enhanced yeast biomass and β-carotene production, which were promoted by41.3% and 52.8% respectively.This paper would offer references to screen the rhodotorula producing β- carotene and viable utilization of β-carotene.