Abstract: Optimization the production of extracellular lipase in culture of Bacillus cereus has been investigated. From 10 samples of diesel fuel polluted soil it was found that 3 samples was positive for the presence of B. cereus. Lipase production by B. cereus isolated from diesel fuel polluted soil was investigated and optimized. The enzyme production was increased with increasing time and maximum enzyme activity was obtained after 72 hrs of incubation. Effect of pH and temperature indicated that, the lipase production was maximum at pH 8.0 (60.2 U/ml) and at 35°C (55.25 U/ml). The lipase production was optimized in shake flask experiments. With a selected carbon sources, maltose (65.5 U/mL) and nitrogen source, peptone (66.25 U/mL) was suitable substrates for accelerating lipase production. The present study indicates that the various factors influenced enzyme production by the bacteria, it appears that the nature of the supplements had significantly influenced on the production.