In the present study, Grass pea protein isolate (GPPI)- Alyssum homolocarpum seed gum (AHSG) complex nanoparticles were formed through two fabrication methods and their physicochemical properties, structure and stability against sodium chloride and different pHs were investigated. Type 1 particles were formed by creating GPPI nanoparticles, and then coating them with AHSG; while Type 2 particles were fabricated through the heat treatment of GPPI-AHSG complexes at 85 °C for 15 min. The preparation methods did not influence the magnitude of electrical charges on biopolymer particles. The particle size analysis revealed that Type 2 particles had lower mean diameter (d = 360.20 nm) compared to Type 1 particles (d = 463.22 nm). Structural properties of Type 1 and Type 2 particles were determined using Fourier transform infrared (FTIR) spectroscopy, X-ray diffractometry (XRD), Differential scanning calorimetry (DSC), Atomic force microscopy (AFM), and transmission electron microscopy (TEM). Hydrogen bonding, electrostatic and hydrophobic interactions were the main driving forces contributed to the formation of both GPPI-AHSG complex particles. Assessments of morphological and structural properties also indicated that both Type 1 and 2 particles had spherical shapes and heat treatment increased the ordered intermolecular structures in biopolymer particles. Type 2 particles had higher denaturation temperature and better pH and salt stability when compared to Type 1 particles. These results indicate that thermal treatment was effective for the fabrication of stable GPPI-AHSG complex nanoparticles.

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