Appropriate selection of the drying temperature is of great importance to achieve desirable film preparation. The present study investigated the impact of drying temperature (40-80 °C) on the physical properties and molecular assembly process of konjac glucomannan/ethyl cellulose/zein blend films through the casting method. Microstructural observations indicated that the drying temperature impacted the particle aggregation in the films, and at the drying temperature of 60 °C, a most even component distribution was suggested with the strongest hydrogen bond interaction according to the FTIR and micro-FTIR analysis. Moreover, at this drying temperature, the blend film reached optimum performance for most physical properties including mechanical properties, surface hydrophobicity, moisture barrier, and water resistance properties. To explain this phenomenon, the molecular assembly process during the KGM/EC/zein film formation was proposed. This contributed to a deepened understanding of the molecular assembly process of complex macromolecules under different drying conditions.