The present study is carried out by isolation, characterization of biosurfactant producing bacteria and their further activities. 12 bacterial species were isolated through serial dilution method from 4 different sites of Lucknow and Unnao district. The serial dilution method was performed in order to get pure and reduced number of bacterial colonies. 6 bacterial species out of 12 were detected of producing biosurfactant activity. These bacterial species were further morphologically and biochemically characterized through Bergey’s manual. These bacterial species were tested for 7 days to oil degradation capacity by agar well diffusion method on small scale and flask method on large scale. The three types of oil samples were used for bio surfactant activities such as vegetable oil, used vegetable oil (cooked oil) and diesel. The best bio surfactant activities were obtained for all these bacterial species against used vegetable oil (cooked oil). The identified cultures were Steptococcus spp., M. luteus, S. aureus, Alcaligens spp., B. cereus, N. flavescens.