The yogurt was prepared by partially replacing cow milk protein in milk powder with soy protein isolate(SPI). The influence of substitution of SPI for milk protein on the properties of yogurt was investigated by measuring the physicochemical properties (pH, titratable acidity, water-holding capacity, and color difference) and textural characteristics. And its influence on the internal structure of yogurt was explored by measuring the potential, particle size, hydrophobicity, microstructure, and protein secondary structure. The results showed that with the increasing of SPI substitution (0% to 50%), the sensory quality of the yogurt was improved, and the structural properties was enhanced. The titratable acidity of the yogurt decreased by 17.66 °T (P<0.05), and the water holding capacity increased to 59.17% (P<0.05). The textural properties were all enhanced (P<0.05), and the particle size and zeta potential increased significantly (P<0.05). In terms of physical structure, as the substitution amount of SPI increased, the soy protein and casein in the yogurt cross-linked with each other to form curd with dense network, which was better than conventional yogurt curd at 30% substitution of SPI, and closer to tofu curd when adding 50% substitution of SPI. In terms of chemical structure, the β-fold content tended to decrease with increase of SPI substitution, while the relative α-helix, β-turn and random curl increased in varying degrees. The addition of SPI significantly enhanced the physicochemical properties and curd structure of the yogurt, and 30% was the optimal substitution amount of SPI.
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