Rapeseed cake serves as a by-product in the oil extraction industry, characterized by its elevated protein content. However, the presence of antinutritional factors limits the utilization of rapeseed cake as a viable protein source. In this study, different doses of γ-irradiation were used to irradiate rapeseed cake and rapeseed protein isolate was extracted through a modified alkaline solution and acid precipitation method from rapeseed cake. The chemical composition and in vivo acute toxicity of rapeseed protein isolate were determined. The protein recovery rate of rapeseed protein isolate was 39.08 ± 3.01% after irradiation, while the content of antinutritional factors was significantly reduced. Moreover, γ-irradiation did not have any experimentally related effects on clinical observations or clinicopathology in mice. Overall, the reduced antinutrients and increased functional properties suggest that the irradiation of rapeseed cake (<9 kGy) could be utilized as a pre-treatment in the development of rapeseed cake-based value-added protein products.
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