High-moisture extrusion (HME) is a promising technology for producing plant-based seafood analogs (PBSAs); however, achieving desirable texture and minimizing off-flavors remain major challenges. In this study, HME-PBSAs were prepared using soybean protein isolate (SPI) and wheat gluten (WG) as the base, with SPI partially replaced by rice protein isolate (RPI), yeast protein (YP), mung bean protein isolate (MPI), pea protein isolate (PPI), or peanut protein isolate (NPI). Results demonstrated that protein source significantly affected rheological behavior, fibrous structure, texture, and flavor retention. The addition of MPI, NPI, and PPI promoted the conversion of β-turns to β-sheets. The NPI-containing extrudate (ENPI) exhibited a relatively high β-sheet content (27%) and the highest fibrous degree (1.59), forming a well-developed layered fibrous network. Texture profile analysis and principal component analysis revealed that ENPI showed textural characteristics comparable to grilled eel, grilled squid, and grilled black scraper. Volatile compound analysis indicated that ENPI contained substantially lower levels of key off-flavor compounds, including hexanal (74.86 μg/kg), nonanal (12.44 μg/kg), 2-heptanone (2778.1 μg/kg), and 2-pentylfuran (5742.21 μg/kg), compared with other formulations. In contrast, extrudates containing YP or RPI exhibited excessive hardness and chewiness, lower fibrous degree, denser microstructures, and higher off-flavor contents, resulting in reduced sensory acceptance. Under the experimental conditions of this study, the incorporation of NPI showed comparatively better performance among the tested protein blends, contributing to improved texture-flavor characteristics in HME-PBSAs. This study provides a scientific basis for protein blend selection and formulation optimization in the development of high-quality PBSAs.
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