Developing prospective plant-animal binary protein systems with desirable nutritional and rheological properties stands as a significant and challenging pursuit within the food industry. Our understanding of the effect of adding salt on the aggregation behavior of food proteins is currently based on single model protein systems, however, this knowledge is rather limited following binary protein systems. Herein, various ionic strength settings are used to mitigate the repulsive forces between pea-cod mixed proteins during the thermal process, which further benefits the construction of a strengthened gel network. Transmission electron microscopy (TEM) and dynamic light scattering (DLS) collectively demonstrated that larger heat-induced protein aggregates were formed, which increased in size with higher ionic strength. In the presence of 2.5 mM CaCl2 and 50 mM NaCl, the disulfide bonds significantly increased from 19.3 to 27.53 and 30.5 μM/g, respectively. Notably, similar aggregation behavior could be found when introducing 2.5 mM CaCl2 or 25 mM NaCl, due to the enhanced aggregation tendency by specific binding of Ca2+ to proteins. With relevance to the strengthened cross-links between protein molecules, salt endowed composite gels with preferable gelling properties, evidenced by increased storage modulus. Additionally, the gelling temperature of mixed proteins decreased below 50 °C at elevated ionic strength. Simultaneously, the proportion of network proteins in composite gels increased remarkably from 82.05 % to 93.61 % and 92.31 % upon adding 5.0 mM CaCl2 and 100 mM NaCl, respectively. The findings provide a valuable foundation for designing economically viable and health-oriented plant-animal binary protein systems.

Isoelectric solubilisation/co-precipitation (ISP) has been proven to be a better method than blending for preparing plant-animal dual-proteins, which can achieve synergies in the functional properties of heterologous proteins. This paper aims to investigate the effect of extraction pH on the functional properties of co-precipitated dual-protein. The basic composition, subunit composition, solubility, surface hydrophobicity, emulsification and gel properties of co-precipitated dual-protein (Co) prepared from pea and grass carp with pH (2.0, 3.0, 9.0, 10.0 and 11.0) were analysed in this study using ISP. The results showed that the functional properties of Co (Co9, Co10, Co11) prepared by alkali extraction were generally better than those prepared by acid extraction (Co2, Co3). Among them, Co10 has the highest vicilin/legumin α + β value and solubility, while having the lowest surface hydrophobicity, making its emulsification and gel properties superior to other extraction pH values. This study provides an important method reference for preparing plant-animal Co with exceptional functional properties.

Although natural emulsifiers often have many drawbacks when used alone, their emulsifying ability and stability can usually be improved unexpectedly when used in combination. In this study, monodisperse emulsions stabilized by combining two natural protein emulsifiers, i.e., whey protein isolate (WPI) and sodium caseinate (SC), in different proportions were prepared using microchannel (MC) emulsification. The influences of temperature, pH, ionic strength, and storage time on the microstructure and stability of the emulsions were examined. Analysis of the microstructure and droplet size distribution revealed that the WPI-, SC-, and mixed protein-stabilized emulsions exhibited uniform droplet distribution. The droplet size and ξ-potential of the MC emulsions stabilized by mixed protein emulsifiers were higher than those of the emulsions stabilized by WPI or SC separately. The emulsions stabilized by the two types of proteins and mixed emulsifiers had better stability under high salt concentrations than the synthetic emulsifier Tween 20. WPI-SC-stabilized emulsions were more resistant to high temperatures (70-90°C) and exhibited excellent stabilization than those stabilized by WPI and SC, which was attributed to the more sufficient coverage provided by the two types of protein emulsifier layers and better protein adsorption at the oil-water interface. These results indicate that WPI-SC is a potential stabilizer for MC emulsion requirements. This study provides a basis for the formulation of monodisperse and stable natural emulsion systems.

Cold-set emulsion gels were fabricated from oil droplets coated by mixed proteins: whey protein and lactoferrin. The impact of protein composition, droplet concentration, pH, and ionic strength on the microstructure, texture, and stability of the cold-set emulsion gels was determined. Protein composition had a major influence on gel strength, with the strongest emulsion gels being formed at an optimized protein composition (0.5 wt% whey protein and 1.5 wt% lactoferrin). The storage modulus of the emulsion gels increased from 149 to 1590 Pa as the droplet concentration increased from 10 to 40 wt%. The gel strength could also be modulated by adjusting pH, with the strongest gels being formed at pH = 6.5, where the net charge on the droplets was neutral. Increasing the ionic strength weakened the electrostatic interactions, which inhibited droplet aggregation and led to a decrease in gel strength. These results may be useful for designing cold-set emulsion gels with rheological properties that can be tailored for specific commercial products.