The production of lipase is aimed from a new fungal source in this study. Fungus was isolated from soil locality Kocaeli. It was determined as Trichoderma citrinoviride by the analysis of 18S rRNA sequence. Various parameters and media components were investigated for production of lipase. Glucose and peptone were found to be most suitable carbon and nitrogen source, respectively. To determine the suitable oil as carbon source, various oils were added to the production medium. Olive oil was found to be the optimal oil for lipase production from Trichoderma citrinoviride. pH 5.5, temperature 30°C and incubation time for 4 days were found to be optimal incubation conditions for lipase production. We also determined lipase yield fromTrichoderma citrinoviride which is produced in molasses medium as an alternative carbon source.