The need for enzymes is growing across a number of industries due to their several advantages, such as their minimal environmental toxicity. Lipases are one of the most valuable biotechnological enzymes. As a result, studies on lipases have become significantly popular in the field of enzymology in recent years. Several attempts have been made to isolate different bacterial isolates that can produce the lipase enzyme. The current study aims to isolate diverse bacterial isolates from different sources: soil contaminated with oil, animal wounds, and contaminated culture media. A qualitative screening for lipase-producing isolates was conducted using tween 80 agar. The results showed that a total of 124 bacterial isolates were obtained from different sources, including 41 isolates that can produce the lipase enzyme. Upon the lipase activity assay, the 20 highest lipase-producing isolates were identified phenotypically. The best potential lipase producers were further identified using 16S rRNA sequencing as Brevibacillus sp. strain HC1 and Brevibacillus sp. strain HS5, with the accession numbers OR048061 and OR048060, respectively. This study highlights the significance of using bacteria as a microbial source for lipase enzyme production for future industrial and biotechnological applications.