Almond protein as Pickering emulsion stabilizer: Impact of microgel fabrication method and pH on emulsion stability

Abstract

We evaluated the ability of almond proteins to produce Pickering emulsions (EM) stabilized by microgels (MG) fabricated by three different methods (heat treatment-HT, crosslinking with transglutaminase-TG or calcium-CA), at two pH levels (pH 3 or 7). Compared to pH 7, acidic pH significantly denatured almond proteins (ellipticity ∼0 mdeg), decreased absolute zeta potential values (10.5 to 18.6 mV at pH 3 and - 24.6 to -32.6 mV at pH 7), and free thiol content (114.64-131.60 μmol SH/g protein at pH 3 and 129.46-148.17 μmol SH/g protein at pH 7 - except in CA-crosslinked microgels, p > 0.05). These changes led to larger microgel sizes (D_3,2pH3: 26.3-39.5 μm vs. D_3,2pH7: 5.9-9.0 μm) with lower polydispersity (Span_pH3: ∼ 1.94 vs. Span_pH7: 2.32, excluding CA-based samples). Consequently, the Turbiscan Stability Index (TSI) was higher in acidic conditions for all emulsions, except for the calcium-containing formulation (EM_CApH3), emphasizing the critical role of calcium binding in maintaining emulsion stability in acidic environments. Microgels prepared via the traditional heat treatment method produced emulsions with intermediate stability (TSI ranging from 3.4 % to 5.1 % at 28 days of storage). Conversely, TG-crosslinked microgels led to unstable emulsions at pH 3, likely due to the lowest zeta potential (+4.2 mV), whereas at pH 7, the greatest stability was attributed to bridging flocculation that created a stable gel-like structure. Indeed, emulsions with lower TSI (EM_CApH3 = 1.8 %, EM_CApH7 = 2.3 % and EM_TGpH7 = 1.0 %, at 28 days of storage) also exhibited higher elastic modulus (G') over frequency sweep, indicating that the strong elastic network was relevant for emulsion stability (up to 28 days). This study, for the first time, demonstrated the production of stable almond-based Pickering emulsions, with properties modulated by the pH and method used to fabricate the microgels.

Keywords: Almond protein; Calcium crosslinking; Pickering emulsion; Transglutaminase.