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Petroleum hydrocarbon oil contaminated soils were excavated from a gas station from Nicosia Cyprus, Turkey and subjected to fungal isolation/analysis. During the fungal essay, Aspergillus nigar, Penicellum notatum and Mucorales were isolated and identified. Aspergillus nigar was selected and cultivated for lipase enzyme production due to its role in lipase production. The produced lipase enzyme was immobilized and the number of lipases recovered after purification was 85.1%. The purified enzymes that were supported by physical adsorption which resulted in high (85%) yield showed high stability than free lipase, encouraged transesterification in this study. Pretreatment evaluation of groundnut powder showed high cellulose yield (82.3% and 87%) and high lignin removal of 75%, 88% respectively for lipase enzyme accessibility and ethanol production. Transesterification involving purified lipases and groundnut shell fermented ethanol yielded Methyl ester which represents the organic biodiesel with a C18 number showing a minimum level of lubricity for friction reduction between surfaces when a car is on motion and has specific gravity 0.95 ul, density 15.5°c, and calorific value of 43.2 MJ. Kg.