Currently, the correlation between how protein structural changes affected the function of high internal phase emulsions (HIPEs) and the regulation of 3D printing performance was still unclear. Herein, this study was aimed to research the relationship between interface properties and stability of soy protein isolate (SPI) at different degrees of oxidation and prepare HIPEs with desirable rheological properties and a solid-like structure to use for 3D printing. The findings of this study suggested that moderate oxidation led to the disruption of the protein's main chain skeleton, resulting in enhancing protein flexibility and surface hydrophobicity. The HIPEs stabilized by this condition owned higher apparent viscosity, over 70% of thixotropic recovery rate, sufficient yield stress (160.8 Pa) and higher G′ and G″. This HIPEs with excellent properties could be the food-grade 3D printing inks to produce samples with high resolution. In addition, this HIPEs exhibited a slow-release effect on free fatty acids (FFA) in the gastrointestinal digestion. Finally, the findings of the correlation analysis indicated that the flexibility and surface hydrophobicity of SPI was a key factor to affect the quality of HIPEs and 3D printing. In conclusion, moderate oxidation could change the structure and interfacial characteristics of SPI, thus endowing HIPEs stabilized by modified SPI with better properties and expanding the field of SPI in the food industry. These findings not only were of great significance for guiding protein production and providing new protein-based 3D printable materials, but also this provided new possibilities for preparing fat substitutes with good appearance.