Lycopene has become one of the most interesting antioxidant compounds, especially in relation to human health. This work describes a genetic strategy to modify the carotenoid biosynthesis pathway to develop a lycopene-overproducing strain. The crgA gene, a negative regulator of carotenogenesis, was disrupted in the Mucor circinelloides strain MU202, which lacks the lycopene cyclase activity and accumulates lycopene instead of beta-carotene. The resultant strain, MU224, demonstrated increased transcriptional levels of the carotenogenesis structural genes carRP and carB compared to the parental strain MU202. As a consequence, strain MU224 accumulated 5 mg/g of dry weight of cells in liquid cultures, a sevenfold increase with respect to the parental strain. Moreover, when lycopene production was examined in a complex enriched medium, biomass increased tenfold compared to that obtained in synthetic minimal medium. In this complex medium, the production rate of lycopene by strain MU224 reached 54 g/1. These results illustrate how a combination of genetic manipulation and optimized culture conditions can be utilized to enhance the production of commercially desirable compounds such as lycopene.