Polythene is a polymerized ethylene monomer which is resistant to degradation accumulates in an environment. Polythene used in everyday life as a packaging material. Release of toxic chemicals from polythene creates negative impacts on environment and life threatening problems to many aquatic and terrestrial organisms. The present study focused on biodegradation of polythene by soil microorganisms .Isolated microorganisms were identified based on biochemical characterization as Staphylococcus aureus and Bacillus cereus. Biodegradation of polythene by isolated microorganisms were tested in vitro in both nutrient and minimal medium. After 30 days incubation period, the biodegradation of polythene was measured in terms of weight. The percentage of weight loss by staphylococcus aureus and Bacillus cereus in nutrient medium was 32.2% and 9% respectively. The percentage of weight loss by staphylococcus aureus and Bacillus cereus in minimal medium was 57.3% and 26% respectively. The results depict that Staphylococcus aureus are found active in degrading polythene than Bacillus cereus.