Moringa oleifera seed rennet as a novel hydrocolloid coagulant in the production of mozzarella cheese: Effects on function, texture, structure, and rheology

This study evaluated the potential of Moringa oleifera seed rennet (MSR) as a novel hydrocolloid coagulant for mozzarella cheese production compared with commercial rennets. Mozzarella cheeses were manufactured using MSR and 3 commercial rennets (calf rennet, CR, papain rennet, PR, rice black mold rennet, MR). MSR-treated cheese (MSRC) showed comparable nutritional value to calf rennet cheese (CRC), with no significant differences in protein and fat content (P > 0.05), while exhibiting superior hardness and elasticity compared with PR- and MR-treated cheeses.Mechanistically, MSR specifically hydrolyzed κ-casein, increasing α-helix content and fluorescence intensity while reducing free sulfhydryl content, indicating strengthened protein secondary and tertiary structures. These changes led to a denser, more stable protein gel network with smaller pore sizes and more uniform casein distribution, as confirmed by microstructure analysis. Rheologically, MSRC exhibited higher G' and G" values, indicating enhanced elastic and viscous gel-like behavior. Correlation analysis revealed strong positive correlations between fat content and hardness (r = 0.995) and between protein content and elasticity (r = 0.951), while α-helix structure was negatively correlated with moisture (r = -0.980), resilience (r = -0.915), and meltability (r = -0.880). These findings demonstrate that MSR is a viable plant-based, sustainable alternative to commercial rennets for mozzarella cheese production.