In this study, gum arabic (GA)-based nanofibres were prepared and characterized in combination with pullulan (PUL) by electrospinning for the first time, and they served as the wall material for Lactobacillus encapsulation. The presence of GA in hybrid solutions resulted in increased conductivity, reduced surface tension and lower viscosity (P < 0.05), and rheological analysis showed pseudoplastic behavior for all blend solutions. Scanning electron microscopy indicated that the addition of GA led to the formation of nanofibres with smaller diameter. Furthermore, X-ray spectra and Fourier transform infrared spectra revealed the miscibility and hydrogen bonding interactions between GA and PUL molecules. Moreover, the GA/PUL hybrid nanofibres exhibited better thermal stability than native GA. The Lactobacillus-loaded GA/PUL 20:80 electrospun nanofibres showed better probiotic survivability (85.38-97.83%) compared to freeze-drying (80.92-89.84%) and retained viability during 28 day storage at 4 degrees C. The application of GA/PUL nanofibres with prebiotic characteristics provides a new approach for the effective protection of probiotics.