Aims: The aim of this study was to cultivate the fungus Aspergillus tubingensis (MO503), which was isolated and identified from Algerian soil using submerged fermentation. The focus was on the production of lipase, achieved through utilizing a minimal medium from agro-food industries. Specifically, the study investigated the potential of three waste sources – olive-pomace, Pistacia lentiscus fruit remains and olive mill wastewater as substrates for enhancing lipase production. Methodology and results: The three aforementioned wastes were chosen to ascertain their value and determine the most cost-effective option. Factorial analysis of variance (ANOVA) was employed to evaluate the waste with a significant effect on lipase production, as this enables determining differences among the various waste sources. Growth monitoring revealed a maximum lipase activity of 1030 ± 0,039 U at pH 5.4 for olive pomace. A series of biochemical purification techniques displayed a visible band on the polyacrylamide gel obtained through SDS-PAGE. Lipolytic activity was evidenced by zymography in the presence of olive oil. The antibacterial activities of purified lipase exhibited a high sensitivity against Gram-negative bacteria compared to Gram-positive bacteria. Conclusion, significance and impact of study In addition to its activity against Gram-negative bacteria, the lipid degradation facilitated by these lipases in olive oil offers promising applications in the textile and therapeutic industries.