Owing to lack of soluble potassium resource in some countries, it will be the competitive alternative to obtain potassium from insoluble K-bearing minerals (e.g. K-feldspar, biotite) in order to satisfy the requirements of potassium fertilizers for the agricultural sustainable development. In this work, seven fungal species including Penicillium oxalicum, P. ochrochloron, P. simplicissimum, P. spinulosum, Aspergillus niger, A. fumigatus, and A. oryzae were investigated for potassium extraction from biotite (9.64% K2O, weight fraction). Experiments were carried out in a shaker (30 °C, 180 rpm) with biotite directly dispersed in the leaching medium or enclosed in a dialysis bag. The concentrations of potassium, silicon and aluminum in the bioleaching solution were measured by Inductive Coupled Plasma (ICP), and K release amounts of biotite by seven fungal species were compared. It was found that all the strains had the abilities to enhance the dissolution of biotite, and P. oxalicum showed the highest K release amount among seven tested strains. Furthermore, the metabolites of fungi in the bioleaching process were analyzed by high performance liquid chromatography (HPLC), and the surface morphologies of biotite before and after bioleaching were characterized by Scanning Electron Microscope (SEM). The mechanism of microbial release of potassium from biotite by Penicillium andAspergillus was discussed.