Hybrid fresh salad cheese analogue: importance of pea to micelle casein protein ratio on structure, texture and protein retention

This study aimed to evaluate the feasibility of producing hybrid salad (Feta-style) cheese analogues by combining pea protein isolate (PPI) and micellar casein isolate (MCI) in varying ratios, and to assess how PPI:MCI ratio influences physicochemical properties and proteomic characteristics of cheese- and whey-analogues. Hybrid cheeses were prepared using Mucor pepsin and chemical acidification, with emulsions standardized to 8.5 % (w/w) protein and supplemented with 15 % (w/w) rapeseed oil. Analyses included colour, texture, pH evolution, yield, and composition. Proteomic profiling was performed to investigate protein retention and enzymatic cleavage during cheese-like formation. The resulting prototypes contained 12.5 to 16.8 % protein and 27.1 to 31.4 % fat which is acceptable for this type of cheese. Salad-cheese-like textures were obtained after draining the whey analogue. Yield increased with higher PPI content, while hardness decreased. Proteomic data revealed secondary cleavage sites of Mucor pepsin on caseins and on pea proteins (legumin B, legumin J, convicilin). Pea proteins were retained in greater proportions than caseins in the final cheese-like matrix. The optimal formulation, 70 % MCI and 30 % PPI, provided a balance between texture and yield, demonstrating the potential of hybrid protein systems for salad-style hybrid cheese production.