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This study focused on the valorization of the date syrup obtained from low-quality date fruits to be used as a low-cost alternative medium for producing single cell oil (SCO) by Rhodotorula glutinis PTCC5256, which could further be converted into biodiesel. The higher C/N ratio of date syrup (C/N 7 led to restricting the formation of cell biomass and enhancing the biosynthesis of SCO. The maximal cell biomass and lipid productivities were obtained 72 mg/L/h and 17 mg/L/h by C/N ratios of 20 and 70, respectively. Although the obtained