Abstract
The aim of this work was to systematically evaluate the effects of Lion's Mane Mushroom powder (LMM, 0-40%) on the physicochemical properties, structural characteristics, and flavor profile of soy protein isolate-based high-moisture meat analogues (HMMAs). Optimal incorporation of 20% LMM significantly enhanced product quality by acting as a secondary phase that inhibited lateral protein aggregation while promoting longitudinal alignment, achieving a peak fibrous degree of 1.54 with dense, ordered fibers confirmed by scanning electron microscopy. Rheological analysis showed that LMM improved viscoelasticity (G' > G″) through β-glucan; however, excessive addition (≥30%) compromised structural integrity due to insoluble dietary fiber disrupting protein network continuity, concurrently reducing thermal stability as denaturation enthalpy ((Δ)H) decreased from 1176.6 to 776.3 J/g. Flavor analysis identified 285 volatile compounds in HMMAs with 20% LMM, including 98 novel compounds, and 101 flavor metabolites were upregulated. The mushroom-characteristic compound 1-octen-3-ol exhibited a marked increase in its Relative Odor Activity Value of 18.04, intensifying mushroom notes. Furthermore, LMM polysaccharides promoted the Maillard reaction, increasing the browning index from 48.77 to 82.07, while β-glucan induced a transition in protein secondary structure from random coil to β-sheet configurations via intramolecular hydrogen bonding. In conclusion, 20% LMM incorporation synergistically improved texture, fibrous structure, and flavor complexity-particularly enhancing mushroom aroma. This research offers valuable insights and a foundation for future research for developing high-quality fungal protein-based meat analogues.