Silkworm (Bombyx moil L.) Pupa protein (SPP) is a high-quality insect protein and is considered a sustainable alternative source for traditional animal food protein. However, the utilization of SPP is limited because of its low solubility and emulsifying ability. In the present study, the synergistic effect of hydration and pulsed ultrasound on the physicochemical properties of SPP and SPP-stabilized Pickering emulsions was evaluated. Pulsed ultrasound changed the particle size of SPP and its conformation. As the pulsed ultrasound increased from 0 s to 5 s, the α-helix and SS contents of SPP decreased, whereas the β-sheet and SH contents increased, which in turn improved its solubility and amphiphilicity. As a result, the SPP treated by a combination of 12 h of hydration and 3 s of ultrasound exhibited a contact angle of 74.95°, hydrophobicity of 904.83, EAI of 6.66 m2 g-1 and ESI of 190.69 min. Compared with the combination of 1 h of hydration and 5 s of ultrasound, the combination of 12 h of hydration and 3 s of ultrasound exerted more soluble and hydrophobic SPP, whereas the EAI and ESI of the samples were higher. Notably, the ultrasound-treated SPP can form a stable gel-like emulsion (oil fraction ranging from 70% to 80%). The combination of hydration and ultrasound can effectively improve the physicochemical characteristics of SPP as well as its emulsion stability. Sufficient hydration is a cost-effective method for facilitating the modification of proteins by ultrasound treatment. © 2024 Society of Chemical Industry.