Fungi have great morphological, physiological, and enzymatic potential that makes them promising organisms to be used in bioremediation processes of unconventional hydrocarbon contaminated soils, such as those found in the Ecuadorian Amazon. Because there are continuous oil spills and drilling cuttings of recalcitrant crude oils in this mega-diverse region with high socio-environmental impact, the present study aimed to isolate and characterize cultivable fungi with high hydrocarbonoclastic potential from asphalt samples at the Pungarayacu pit in the Napo Province, Ecuador. 6 fungal strains were isolated and molecularly identified by PCR amplification and sequencing of the ITS2 region (barcode) belonging to the following genera and species: Fusarium croci, Coniochaeta hoffmannii, Xenoacremonium falcatum, Trichoderma yunnanense, Penicillium amphiboloria and, Neocosmospora rubicola. Once the isolated strains had been identified, a Genome Mining study was carried out to identify possible genes and proteins associated with bioremediation processes in the orange yeast fungus genome Coniochaeta hoffmannii, (https://youtu.be/0vy2NQWPTEQ) focusing the studies on laccase enzymes. In Silico results showed the hypothetical enzyme Lac3 possesses essential characteristics to be considered biological catalysts in environmental mycoremediation processes. The preliminary results obtained in the present study provide a solid foundation to continue exploring the potential of indigenous fungal strains isolated from natural asphalt outcrops in the Amazon rainforest for the bioremediation of oil-polluted soils and the restoration of impacted natural ecosystems