Uranium inventory in Boeun aquifer is facing the artificial reservoir that intended for supplying water to nearby cities (40-70 m apart) where, toxic radionuclides might mobile and enter the reservoir. In order to understand U mobility in the system, groundwater and fracture filling materials (FFMs) were analyzed for microbial signatures, C, O, Fe, S and U-series isotopes. The δ18O-H2O and 14C signatures suggested groundwater was originated from upland recharges dominantly and not affected by mixing with the surface water. However, the 234U/238U activity ratios (ARs) and 230Th/234U ARs in FFMs ranged from 0.93 to 1.67 and from 0.22 to 1.97, respectively, indicating that U was mobile along the fractures. In shallow FFMs, the U accumulations (~157 mg/kg) were found with Fe enrichments (~226798 mg/kg) and anomalies of δ56Fe and δ57Fe, implied U mobility in shallow depths was associated with Fe-rich environment. Also, in the shallow depths, Fe-oxidizers, Gallionella was prevailing in groundwater while Acidovorax was abundant near U ore depth. The Fe-rich environment in shallow depths was formed by pyrite dissolution, demonstrated using δ34S-SO4 and δ18O-SO4 distribution. Conclusively, the Fe-rich aquifer was capable of immobilizing the dissolved U through biotic and abiotic processes, without significant discharge into the nearby reservoir.