Zein is a hydrophobic protein produced from maize and has great potential in a number of industrial applications, such as food, food coating and food packaging. To obtain suitable electrospinning conditions for thinner and uniform zein nanofiber mats, a series of experiments was conducted on various volume ratios (v/v) of ethanol/water solutions with different zein concentrations. The prepared zein nanofiber mats were characterized by field emission scanning electron microscopy and contact angle measurements. Uniform zein fibers with a average diameter in the nanometer scale (300~500 nm) could be prepared from 30 wt.% zein in 7/3 (v/v) ethanol/water solutions.