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 (_ΔH) 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

This work explores the impact of ergosterol (ERG) addition (0%, 0.5%, 1.0%, 1.5%, and 2.0%) on the physicochemical properties, conformational changes, and digestive characteristics of soy protein isolate (SPI) and wheat gluten (WG) processed by high-moisture extrusion. The results demonstrated that the incorporation of ERG significantly reduced the apparent viscosity and dynamic moduli of the feedstock system, enhancing melt fluidity and consequently reducing extrusion torque, die pressure, and specific mechanical energy. An appropriate amount of ERG (1.0%) effectively facilitated the development of a distinct fibrous morphology, increased the fibrous degree, lightened the color, and softened the texture. However, excessive addition weakened the fibrous structure due to excessively high fluidity. ERG influenced protein aggregation behavior through hydrophobic interactions, reduced thermal stability, and induced a transition in secondary structure from β-turns to α-helices. The in vitro digestibility initially decreased and then increased, with the lowest value observed at 1.0% ERG. This study indicates that ERG can elevate the performance and value of extruded products by modulating protein structure and rheological behavior, providing a theoretical basis for its application in plant-based meat analogue products.

Physalis pubescens L. is a seasonal fruit with high nutritional value but a short shelf life that limits its processing and utilization. This study integrated metagenomics and metabolomics to investigate the comparative effects of Lactiplantibacillus plantarum fermentation on volatile flavor metabolites and microbial community composition of P. pubescens by comparing initial (0 h) and post-fermentation (24 h) states. After 24 h of fermentation, 1316 volatile compounds were putatively identified by GC-MS, with 592 metabolites significantly changed and 501 upregulated and 91 downregulated. Key flavor compounds that impart citrus, floral, fruity, and rose notes including D-limonene, geraniol, D-carvone, and phenylethyl alcohol were markedly increased. Metagenomic analysis revealed that L. plantarum rapidly dominated the microbial community (relative abundance surged from <0.05% to ~72%) while effectively suppressing potential spoilage bacteria such as Escherichia coli. Functional gene annotation demonstrated significant enrichment of amino acid, carbohydrate, and fatty acid metabolism pathways, with key enzyme genes (L-lactate dehydrogenase, pyruvate oxidase, acetyl-CoA carboxylase) predominantly assigned to L. plantarum, suggesting their potential contribution to the generation of organic acids, ethanol, and esters. Spearman correlation analysis indicated that Lactobacillaceae genera were significantly positively correlated with terpenoids, phenols, alcohols, and aldehydes. This study provides the first metagenomics-metabolomics insight into the microbial and molecular mechanisms associated with flavor formation in LAB-fermented P. pubescens, offering a theoretical foundation for developing stable and controllable fermented fruit products.