The bacterial strain having the ability of minerals bioleaching and desulphurization of coal has been isolated from coal mine drainage and hot spring water collected from the district Harnai, Balochistan. The recent work involves sample collection, growth culture, purification, and confirmation of bioleaching activity present in isolated strain with suitable evidence. The isolated Thiobacillus ferrooxidans bacteria showed the ability to decrease the pH of the medium during its growth along with the conversion of Fe (II) to Fe (III) state. The optimization of various parameters that affect the growth culture has been investigated suggesting the use of isolated bacterial strain for the production of sulphuric acid as well as for minerals leaching from respective ores and coal desulphurization.Keywords: T. ferrooxidans, Growth culture, Isolation, Harnai, Balochistan.Citation: Hafeez, I., Mushtaq, A., Aamir, M., Amin, A., Akram, A., 2020. Isolation and Growth Culture of Thiobacillus ferrooxidans from Coal Mine Drainage of Harnai, Balochistan. PSM Microbiol., 5(1): 1-6.