A novel fluorine-containing copolymer surfactant was synthesized via free radical copolymerization from maleic anhydride and rapeseed oil firstly, and then followed by an esterification reaction with dodecafluoro heptanol. The reaction progresses were monitored on-line by FTIR and the copolymerization was characterized by molecular weight (GPC) analysis. Simultaneously, the surface tension and critical micelle concentration (CMC) together with the biodegradability of the fluorine-containing copolymer surfactant were investigated in detail. The results indicate that this fluorine-containing surfactant with Mw of 66000 g/ mol and Mn of 38800 g/mol shows a strong surface tension lowering ability, the CMC is 0.1 g/L, the surface tension value at the CMC (γCMC) is 22.5 mN m−1 and less than the corresponding copolymeric hydrocarbon surfactant; the evaluation of biodegradability indicates that the biological respiration curve of the fluorine-containing copolymer surfactant with concentration of 1000 mg/L is above the endogenous respiration curve; the BOD5/COD value is higher than 0.45; the COD and TOC removal ratios after 5 days of biodegradation reach 91.0 and 89.4%, respectively, showing good biodegradability and environmental-friendly feature.