In this work aimed to optimize the culture condition of the fungal strain Thermothelomyces thermophiles XJF-9 to increase the growth and production of pharmacological active secondary metabolites. The potent fungus is subjected to optimization in various culture medium and incubation time. The optimized culture medium was Potato Dextrose Broth (PDB) using dextrose as carbon source and incubation time 14 days for the maximum growth and production of bioactive natural compounds by T. thermophiles XJF-9. Moreover, secondary metabolites of crude ethyl acetate extract were investigation melanin content assay and tyrosinase activity. These secondary metabolites are effectively utilized in medicinal and pharmaceutical industry.