Impact of cell disruption technologies on the structure, functionality, and digestibility of Cordyceps militaris protein isolates

Cell disruption is essential for extracting intracellular proteins from fungal biomass. However, its effects on the structural and functional properties of proteins remain insufficiently understood. This study used Cordyceps militaris as a model to evaluate three cell disruption techniques—high-pressure homogenization (HPH), ultrasonication (US), and cellulase hydrolysis—for extracting C. militaris protein isolates (CMPI). All methods significantly improved protein extraction compared to alkaline extraction alone, with HPH yielding the highest rate (70.75 %, p  < 0.05). Structural analysis revealed a decrease in α-helix and β-sheet content, along with increased β-turns, random coils, surface hydrophobicity, and modifications in disulfide–sulfhydryl balance. CMPI-HPH and CMPI-US showed superior water-holding capacities (6.23 and 5.95 g/g) and oil-holding capacities (9.97 and 7.70 g/g), as well as enhanced foaming (69.57 % and 65.22 %) and emulsifying activities (32.07 and 31.13 m 2 /g). CMPI-HPH also exhibited the highest foaming stability (87.50 %). In vitro digestibility was significantly greater in CMPI-US (80.05 %) and CMPI-HPH (71.71 %) than in the control ( p  < 0.05). These results demonstrate that cell disruption, especially HPH, effectively improves protein extraction efficiency, structural flexibility, and techno-functional properties, offering new potential for the sustainable application of fungal proteins in food systems.