The creation of plant-based meat analogs has gained interest recently due to the need to produce a more sustainable and environmentally friendly food supply. Consumers expect meat analogs to have structural and textural attributes that closely mimic those of animal meat, which has led to the need for novel processing technologies to create these kinds of fibrous products. Freeze-structuring has recently gained interest because it is a mild processing operation capable of creating aligned protein-rich fibrous structures in composite biopolymer matrices that resemble the fibers found in muscle foods. However, the time required to produce aligned structures using the current generation of freeze-structuring methods is too long for commercial applications. In this study, cold-triggered release of cationic calcium ions (Ca2+) from the internal aqueous phase of water-in-oil-in-water (W/O/W) emulsions was used to promote crosslinking of anionic biopolymers (alginate and potato protein) dissolved in the external aqueous phase of these emulsions. Initially, these composite materials were unidirectionally frozen, which led to the formation of long thin ice crystals within the composite biopolymer matrix. As a result, aligned biopolymer-rich fibers were formed between the ice crystals. The freezing-thawing process also promoted disruption of the W/O droplets in the double emulsions, leading to release of cationic Ca2+ ions and the crosslinking of the anionic alginate molecules within the biopolymer-rich fibers. This process allowed us to rapidly fix the fibrous structures formed after freeze-structuring. The textural attributes of the biopolymer composites could be further improved by heat-setting the potato proteins through controlled thermal denaturation and aggregation. Protein concentrations (10 % or 15 %), pH conditions (pH 7.0 or 5.7), and freezing speed (slow or fast) influenced the texture and structural anisotropy of the composite biopolymer materials. Microstructural analysis using confocal fluorescence microscopy showed that the fibers formed were highly aligned at the lower protein concentration (10 %), lower pH value (pH 5.7), and slower freezing speed. The triggered release system developed in this study could be used to rapidly produce freeze-aligned structures, which may be useful for improving the structure and texture of plant-based meat analogs.
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