The "Zeeman effect" offers unique opportunities for magnetic manipulation of the spin degree of freedom (DOF). Recently, valley Zeeman splitting, referring to the lifting of valley degeneracy, has been demonstrated in two-dimensional transition metal dichalcogenides (TMDs) at liquid helium temperature. However, to realize the practical applications of valley pseudospins, the valley DOF must be controllable by a magnetic field at room temperature, which remains a significant challenge. Magnetic doping in TMDs can enhance the Zeeman splitting, however, to achieve this experimentally is not easy. Here, we report unambiguous magnetic manipulation of valley Zeeman splitting at 300 K (g eff = -6.4) and 10 K (g eff = -11) in a CVD-grown Fe-doped MoS 2 monolayer; the effective Landé g eff factor can be tuned to -20.7 by increasing the Fe dopant concentration, which represents an approximately fivefold enhancement as compared to undoped MoS 2 .Our measurements and calculations reveal that the enhanced splitting and g eff factors are due to the Heisenberg exchange interaction of the localized magnetic moments (Fe 3d electrons) with MoS 2 through the d-orbital hybridization.