This study was to screen efficient biosurfactant producing strains from waste edible oils (WEOs). Two strains Y5 and Y17 were screened out by steps of preliminary blood plate screening and secondary flask fermentation screening. The surface tension of the fermentation broth of the two strains diluted by 100 times was lower than 40 mN/m. Preliminary characterization of fermentation products was conducted, which was found to be rhamnolipid ultimately. Y5 strain produced only one kind of glycolipids while the Y17 produced at least three kinds of homologues of glycolipids. Both Y5 and Y17 were identified as Pseudomonas aeruginosa strains. This study gave a simple and efficient protocol to screen out biosurfactant producing strains and would pave the way for the production of biosurfactant from WEOs.