Ninety-six isolates from different sources had been isolated and their lipolytic activities investigated using phenol red agar plates. Examine the properties of the lipase enzyme that was separated from the most active isolate of bacteria (MC1) on cow's milk fat. The enzyme showed activity (23.06 U/ml) over an incubation period on 24 h. The biochemical and morphological identification of the bacterial isolate revealed that it was Bacillus sp. The stability of MC1 lipase under varied temperature conditions was enhanced by studying its characterization; it exhibited a maximal activity of 36.17 U/ml at 40 °C and remained stable up to 70 °C with 17% of its relative activity. The lipase enzyme was shown to be more stable at different pH levels (ranging from 5 to 10); at pH 7, the maximal lipase activity was 29.57 U/ml. Addition of metal ion like Ca²⁺ enhancing the lipase activity. It is favored in biological and industrial processes for this reason.