In this study, biodegradable zein protein obtained from maize was used as a novel material to prepare a membrane for oxygen/nitrogen separation. Scanning electron microscopy (SEM), thermogravimetric analysis (TGA), tensile test, X-ray diffraction (XRD) spectroscopy, differential scanning calorimetry (DSC), biodegradability test, and attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy were conducted for characterizing the zein membrane, and membrane properties was compared with that of the polymeric membranes reported in the literature. The developed zein membrane showed a significantly high thermal stability. The mechanical properties of the zein membrane were higher or comparable to those of the previously reported dense membranes. In addition, biodegradation in compost was significantly higher than the other membranes reported in the literature. The developed zein membrane provided a significantly high oxygen/nitrogen selectivity at 2 bar, which was superior to most of the previously reported data on the neat biodegradable polymers with dense structure. The permeability of oxygen and nitrogen at 2