Hydrogen fuel is a promising alternative to fossil fuel. Therefore, efficient hydrogen production is crucial to elucidate the distinctive reactivities of metal hydride species, the intermediates formed during hydrogen activation/evolution in the presence of organometallic catalysts. This study uses density functional theory (DFT) to investigate the isomerizations and reactivities of three nickel-iron (NiFe) hydride isomers synthesized by mimicking the active center of NiFe hydrogenase. Hydride transfer within these complexes, rather than a chemical reaction between the complexes, converts the three hydrides internally. Their reactivities, including their electron-transfer, hydride-transfer and proton-transfer reactions, are investigated. The bridging hydride complex exhibits a higher energy level for the highest occupied molecular orbital (HOMO) than the terminal hydride during the electron-transfer reaction. This energy level indicates that the bridging hydride is more easily oxidized and is more susceptible to electron transfer than the terminal hydride. Regarding the hydride-transfer reaction between the NiFe hydride complex and methylene blue, the terminal hydrides exhibit larger hydricity and lower reaction barriers than the bridging hydride complexes. The results of energy decomposition analysis indicate that the structural deformation energy of the terminal hydride in the transition state is smaller than that of the bridging hydride complex, which lowers the reaction barrier of hydride transfer in the terminal hydride. To produce hydrogen, the rate-determining step is represented by the protonation of the hydride, and the terminal hydrides are thermodynamically and kinetically superior to the bridging ones. The differences in the reactivities of the hydride isomers ensure the precise control of hydrogen, and the theoretical calculations can be applied to design catalysts for hydrogen activation/production.