The effect of electrostatic field-assisted freezing on the gluten protein rheological properties and thermal properties were investigated in this paper. And the molecular structure, the contents of disulfide bonds, ionic bonds, hydrogen bonds, and hydrophobic of the gluten protein was analyzed to explore the improvement effect of electrostatic field assisted freezing on the quality of gluten protein. The results showed that electrostatic field assisted freezing improved the viscoelasticity, the denaturation temperature (Tp) and denaturation enthalpy (?H) of gluten protein compared with the control (traditional freezing method). The analysis of chemical interaction revealed that the content of hydrogen bond decreased first and then increased, while the changes of ionic bond, hydrogen bond and hydrophobic interaction had an opposite tendency. The imaging results of the laser confocal micro-Raman spectrometer indicated that the distribution of secondary structure and the configurations of disulfide bond in the gluten protein became more uniform, and the content of ?-helix and g-g-g configuration increased with the 600 V voltage electrostatic field treatment. SEM image analysis showed that the network structure of gluten protein has smaller pores and distributed evenly under the 300 V and 600 V voltage electrostatic fields.