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Introduction: Microbial lipids attract attention of many researchers due to their therapeutic effects. The goal of this study is the production and optimization of lipids and fatty acids in Mortierella vinacea by applying different media to achieve invaluable fatty acids in pharmaceutical and food industry. Materials and methods: Mortierella vinacea was cultured on potato dextrose agar. Then the spores were inoculated to the production medium. After 72 hours, the lipids were extracted and they were analyzed by gas chromatography. To optimize lipid and important fatty acids production in medium, various carbon and nitrogen sources were substituted with glucose and yeast extract respectively. Results: The effect of some carbon and nitrogen sources on biomass, lipid and fatty acids production were assayed. The highest level of lipid production was in a medium which contains lactose and yeast extract (26.66%). Linoleic acid was only produced in presence of lactose and yeast extract (25.7%). While, M. vinacea yielded the highest level of linoleic acid (52.76%) in a medium containing peptone, linolenic acid was achieved only in presence of lactose and triptone. Discussion and conclusion: In this study, lactose as a carbon source was the most effective one in the production of lipids. In addition, linoleic acid was produced in presence of lactose, so lactose was selected as the best carbon source. Peptone and triptone as a nitrogen source were chosen for the production of linoleic acid and linolenic acid in M. vinacea respectively. All of these findings reveal that Mortierella strain is a potential candidate for enhancement of linoleic acid and linolenic acid production. Furthermore, this simple media can be used in production of linoleic acid and linolenic acid for industrial goals in large scales. [ABSTRACT FROM AUTHOR]