In the present world of industrial revolution, spills of petroleum products­­­­ and oil are one of the major sources of the contamination in the ecosystem and cause serious health hazards for humans and livestock. Therefore, the removal of these pollutants has become necessity of the time. Applications of new techniques such as bioremediation and biodegradation, over conventional methods are much more promising for safe and sustainable environment. Aim-The aim of this study is to isolate and characterize microbial strains from effluent of Mathura Refinery – Indian Oil Corporation Limited, the microscopic and biochemical studies and their potential of bioremediation. Bacteria were isolated from Mathura refinery IOCL, Based on the biochemical studies six morphologically distinct bacterial strains with promising bioremediation role were identified. To check the degradation of crude oil sample by bacterial isolates Bushnallhass media and separation funnel method were performed. Six bacteria were isolates from Mathura refinery effluentsand all six bacteria have ability to degrade the crude oil in different range. Maximum potential to degrade the crude oil were identified in Pseudomonas. This study might be important in an important step towards bioremediation techniques.