The moderate thermophilic mix culture bacteria were used to depyritize the Illinois coal of varying particle sizes (-100 μm, 100-200 μm, +200 μm). Mineral libration analysis showed the presence of pyrite along with other minerals in coal. Microbial depyritization of coal was carried out in stirred tank batch reactors in presence of an iron-free 9K medium. The results indicate that microbial depyritization of coal using moderate thermophiles is an efficient process. Moreover, particle size of coal is an important parameter which affects the efficiency of microbial depyritization process. At the end of the experiment, a maximum of 75% pyrite and 66% of pyritic sulphur were removed from the median particle size. The XRD analysis showed the absence of pyrite mineral in the treated coal sample. A good mass balance was also obtained with net loss of mass ranging from 5-9% showing the feasibility of the process for large scale applications.