Ultrasonication modifies the structural, thermal and functional properties of pumpkin seed protein isolate (PSPI).

Protein isolates from pumpkin seeds were prepared and then treated with high-intensity ultrasound (HIUS) using a probe-based method. The impact of ultrasonication on the physicochemical, molecular, and thermal properties of these isolates were analyzed and compared to untreated controls. Results showed significant improvements (p ≤ 0.05) in color (L*, a*, b* values), solubility, emulsification capacity, and stability, as well as a reduction in molecular weight, indicating enhanced functionality of the pumpkin seed protein isolates (PSPIs) after HIUS treatment. However, HIUS treatment decreased the denaturation temperature (Td), denaturation enthalpy (ΔH), thermal stability, and particle size of the isolates. With treatment durations ranging from 5 to 20 min, T dropped from 67.31 °C to 56.38 °C, and ΔH declined from 45.78 to 35.43 J/g, likely due to structural and conformational modifications from ultrasonic-induced molecular bond disruptions. The greatest reduction in particle size, from 117.46 μm to 85.26 μm, was observed after 20 min of ultrasonication. X-ray diffraction (XRD) analysis showed two distinct diffraction peaks at 2θ = 10° and 2θ = 20°, indicating altered crystallite sizes post-ultrasound treatment. Ultrasonication induced structural and conformational changes in the pumpkin seed protein isolates, as confirmed by SDS-PAGE and weight loss analyses. Alterations in the SDS-PAGE profile and reduced weight loss were associated with improved solubility and enhanced thermal and functional properties in the treated pumpkin seed protein isolates. This emphasizes the potential of PSPI to increase their value-added potential through ultrasonication.