The persistent challenge of membrane contamination, primarily stemming from the extensive usage of non-decomposable materials, stands as a significant barrier limiting the application of microporous membranes in fields like food packaging and biomaterials. This study focused on utilizing indium tin oxide (ITO) conductive glass coated with polylactic acid (PLA) as the electrode. Through electrical induction, zein was prompted to self-assemble, resulting in the creation of porous zein bilayer microporous membranes (PLA/Zein-BMMs). These membranes underwent comprehensive characterization of their physicochemical properties and structure. The outcomes revealed that an electric field voltage of 100 V, an electrode distance of 25 mm, a current density of 15 A/m2, and a protein concentration of 0.05 g/mL, were most suitable for generating microporous membranes featuring a pore diameter of 2 μm and a thickness of 8 μm. During this stage, the microporous membrane exhibited an interconnected three-dimensional network structure, achieving a tensile strength of 49 MPa. Simultaneously, it showcased a surface porosity of 8.15%, a water contact angle measuring 74.28°, and a water vapor transmission rate of 3.2 × 10-3 g/m2•h•KPa. These attributes confer significant application potential in the realms of food packaging and preservation. Additionally, owing to its porous structure and hydrophilic nature, it exhibited favorable biocompatibility when cultured with mouse myoblasts (C2C12), indicating its promising value in the field of biomaterials as well.