This study explores the fabrication of nanofibers using different types of gelatins, including bovine, porcine, and fish gelatins. The gelatins exhibited distinct molecular weights and apparent viscosity values, leading to different entanglement behavior and nanofiber production. The electrospinning technique produced nanofibers with diameters from 47 to 274 nm. The electrospinning process induced conformational changes, reducing the overall crystallinity of the gelatin samples. However, porcine gelatin nanofibers exhibited enhanced molecular ordering. These findings highlight the potential of different gelatin types to produce nanofibers with distinct physicochemical properties. Overall, this study sheds light on the relationship between gelatin properties, electrospinning process conditions, and the resulting nanofiber characteristics, providing insights for tailored applications in various fields.