The effect of more ZnO molecule in tetragonal structure of MOF-5 than cubic structure on the gas permeation properties of T-MOF-5/polyetherimide mixed matrix membranes was investigated. T-MOF-5 was first successfully synthesized and carefully characterized by XRD, FTIR, SEM and N2 adsorption technique at 77 K. Novel T-MOF-5/PEI MMMs were prepared using solution casting method and characterized by FTIR and SEM. The SEM pictures of the MMMs showed that T-MOF-5 nanocrystals changed the morphology of PEI and exhibited acceptable contacts between the filler particles and the polymer chains. Gas permeation properties of these membranes with different TMOF-5 contents were studied for pure H2, CO2, CH4 and N2 gases. Permeation measurement showed that the all gases’ permeability, diffusivity and solubility were increased with T-MOF-5 loading. H2 permeability and the ideal selectivity of H2/CO2 and H2/CH4 in MMM with 25 wt% loading of T-MOF-5 nanocrystals were increased. This behavior was attributed to more ZnO molecule in T-MOF-5 structure. The experimental gas permeations through T-MOF-5/PEI nanocomposite with different filler loadings were fitted on Higuchi model. Good agreement between the experimental data and the predicted gas permeability was obtained.