Abstract:Functional proteins such as food enzymes, flavor-enhancing proteins, pigment proteins and milk proteins have been widely used in food processing industry. By improving food texture, enhancing flavor, imparting color, or boosting nutritional value, these proteins increased the added value of food products. However, general issues for the production of these functional food proteins were low synthesis efficiency and higher costs. Microbial cell factories constructed by synthetic biology technologies offered a green and efficient biomanufacturing approach, enabling the high-yield synthesis of various functional food proteins from low-cost biomass feedstocks. Focusing on four representative categories of functional food proteins, food enzymes, flavor-enhancing proteins, pigment proteins, and milk proteins, this review provided a detailed overview of their specific applications and contributions in improving food texture, flavor, appearance, and nutrition. Based on this, recent research progress, key technological breakthroughs, and the current development status of functional food protein biomanufacturing driven by synthetic biology and other advanced technologies were systematically summarized. Finally, the future opportunities and challenges in the field of functional food protein biomanufacturing were reviewed. This review aimed to deepen understanding of biomanufacturing technologies for functional food proteins and to provide valuable insights for fostering continued innovation and industrialization in this domain.

Abstract:The green off-flavor was an important aroma characteristic in wine and played an important role in the wine quality. The widely present green off-flavor substances in grapes and wines could be primarily classified into 3 categories:methoxypyrazines; green leaf volatiles mainly composed of C6/C9 aldehydes, alcohols, and esters generated through the lipoxygenase pathway from unsaturated fatty acids; and other green off-flavor substances represented by certain sulfur compounds. The regulation of green off-flavor substances was of great significance due to their significant impact on the aromatic characteristics of wine. Currently, the regulation could be carried out from grapes and wine making and storage process. Due to the significant relevance between the content of green off-flavor substances in grapes and the factors such as the maturity and environmental parameters, the adjustment of harvesting time, nutrient regulation(foliar nitrogen application during reraison), light exposure management, water control, pest and disease management and other vineyard measures could achieve the effective regulation of the content of the green off-flavor substances during vineyard management. Apart from the grape material, numerous factors in the whole fermentation, packaging and storage process could also influence the green off-flavor substances. However, different ways of regulation could affect other aspects such as flavor compounds, sugar and acid contents of grape and wine quality. While regulating the content of the green off-flavor substances, the aromatic integrity and matrix effect were also important factors that could affect the whole aromatic quality. This work aimed to clarify the substantial basis of the green off-flavor and summarize the major and potential regulation ways to provide reference for further research and industry application.

Abstract:Chinese whisky gradually established a regionally distinctive flavor profile by innovating in local raw materials, yeast strains, oak barrel selection, and storage techniques while inheriting traditional craftsmanship. The current research on the flavor of Chinese whisky was systematically summarized, focusing on the composition and formation mechanisms of flavor substances, including the dynamic changes of esters, alcohols, acids, aldehydes, and other compounds during fermentation, distillation, and aging. The impacts of oak barrel types, raw material selection, aging processes, yeast strains, and distillation techniques on flavor characteristics were further explored. The results indicated that esters and alcohols were core flavor components:esters significantly enhanced aroma complexity with prolonged aging, while alcohols exhibited regional diversity due to variations in raw materials and processes. Oak barrels contributed characteristic flavors such as vanillin and syringaldehyde through lignin degradation, while the application of domestic Mongolian oak barrels accentuated distinctive oriental sandalwood characteristics. Diversified grain selections and synergistic yeast metabolism provided a foundation for flavor innovation. Additionally, distillation methods markedly influenced the retention and separation of volatile compounds, directly shaping the final flavor layers. This review aimed to consolidate current research findings, offering scientific insights and technical references for optimizing the flavor quality of domestic whisky, promoting standardized production, and enhancing international competitiveness. It also highlighted future priorities, including interdisciplinary collaboration, the establishment of a localized flavor database, and the exploration of innovative aging technologies and raw material adaptability studies.

Abstract:In order to study the effect of loquat leaf extract (LLE) combined with exercise (Ex) on lipid metabolism, mice were divided into normal diet group, high-fat diet group, Ex intervention group, LLE intervention group and LLE-Ex intervention group. After 13 weeks, the body weight changes, organ indices, serum and organ lipid levels, oxidative stress status, protein expression of lipid metabolism and oxidative stress signaling pathways, and serum metabolomic differences of mice were analyzed. The results showed that LLE-Ex could significantly inhibit the body weight gain, lower the adipose tissue proportion in the body, which decreased the index of liver by 8.1%, that of epididymal fat by 22.9%, and that of abdominal fat by 54.0%. It significantly regulated the serum TG, HDL-C and LDL-C levels, and decreased the levels of serum TG and LDL-C by 47.1% and 28.5% respectively, and increased HDL-C levels by 26.2%. It also significantly prevented the deposition of hepatic free fatty acid (FFA) and muscular TG, TC, FFA, and decreased the levels of hepatic FFA and muscular TG, TC and FFA by 41.0%, 13.4%, 36.4%, and 34.9%, respectively. It also improved the level of oxidative stress status in skeletal muscle by decreasing MDA levels at skeletal muscle by 19.5% and increasing CAT activity by 11.5%. The observed effects of LLE-Ex were more comprehensive and marked than LLE or Ex alone, and probably achieved by modulating the AMPK and Nrf2-HO1 signaling pathways and affecting D-glutamine and D-glutamate metabolism, alanine, aspartate and glutamate metabolism and sphingolipid metabolism.

Abstract:Medium chain triglyceride (MCT) is dietary fat naturally existed in palm oil and breast milk. As a novel functional ingredient, it can be used in the production of formula food for special medical purpose. However, the specific target for MCT regulating lipid metabolism is still unclear. Based on network pharmacology analysis and rat model validation, the potential role and target of MCT for regulating lipid metabolism were explored. Animal model demonstrated that no hepatorenal toxicity in rat was observed after nutritional intervention of MCT, indicating its good edible safety. And MCT nutritional intervention down-regulated the levels of serum triglyceride, total cholesterol and low-density lipoprotein, and inhibited the accumulation of adipose tissue of the epididymis in rat. Network pharmacology analysis showed that there were 27 overlapping targets between MCT metabolites (octanoic acid and decanoic acid) and lipid metabolism-related genes, and potentially involved in regulation of peroxisome proliferator activated receptor lipid metabolism-related pathways and fatty acid binding and transport. Western blot assay also indicated that MCT could down-regulate the expression of fatty acid-binding protein 4 (FABP4) of the small intestine in a dose-dependent manner. Results indicated that the MCT nutritional intervention could down-regulate the expression of FABP4, subsequently affecting the absorption and transport of fatty acid, reducing the level of serum lipids and alleviating fat accumulation, thereby regulating the lipid metabolism. The purpose of this study was to clarify the role of MCT in regulating lipid metabolism and its potential targets, and to provide a theoretical basis for the application of MCT in lipid metabolism regulation.

Abstract:i>Lactiplantibacillus plantarum is a probiotic with multiple health benefits. To investigate its effects against alcoholic liver injury, 16 strains of Lactiplantibacillus plantarum were isolated from traditional fermented foods. Their ethanol degradation capacity and effects on alcohol dehydrogenase and acetaldehyde dehydrogenase activities were evaluated, and 8 strains that demonstrated high alcohol metabolism rates and reduced hepatic oxidative stress levels were preliminarily screened. Subsequent assessment of antioxidant capacity, acid resistance, and bile salt tolerance of these 8 strains was carried out, and an optimal strain, Lactiplantibacillus plantarum FS5-5 was identified. An alcoholic liver injury mouse model was established to investigate the effects of Lactiplantibacillus plantarum 5-5. Results showed that FS5-5 exhibited a strong alcohol metabolism capability with alcohol dehydrogenase and acetaldehyde dehydrogenase activities reaching 35.99U/mL and 162.11U/mL, respectively. Meanwhile, high-dose FS5-5 significantly prolonged drunkenness duration, shortened sobering time, and reduced serum ethanol concentration (P<0.05). Furthermore, the high-dose FS5-5 group (1×10⁹CFU/mL) notably alleviated pathological liver damage, decreased hepatic malondialdehyde and triglyceride levels, increased glutathione content, reduced serum transaminases and inflammatory factor levels, enhanced antioxidant enzyme activity, effectively mitigated hepatic oxidative stress (superoxide dismutase level) and inflammatory injury (IL-6, IL-1β, TNF-α levels), and lowered total cholesterol content. Mechanistic studies demonstrated that this strain exerted hepatoprotective effects against alcoholic liver injury by regulating hepatic gene Nrf2, Cyp2e1, NF-κB, and Tlr4 expression, reducing pro-inflammatory cytokine levels, and alleviating oxidative stress and lipid accumulation. This research provided theoretical basis for developing functional foods with auxiliary protective effects against alcoholic liver injury.

Abstract:To explore the effects of covalent modification of locust bean gum (LBG) on the physicochemical properties, structural characteristics and Pickering emulsion stability of pine kernel protein (PKP), infrared spectroscopy and fluorescence spectroscopy were used to analyze the structural differences between conjugates and physical mixtures. The mechanism of interaction between PKP and LBG at different mass ratios was revealed by measuring particle size, Zeta-potential and surface hydrophobicity. Pickering emulsions were prepared using PKP-LBG conjugates as emulsifiers, and the effects of oil phase volume fraction on emulsion properties and stability were investigated. The results showed that compared with the physical mixtures of PKP and LBG, the content of α-helix and β-sheet in the conjugates decreased, and the maximum fluorescence emission wavelength redshifted, indicating that covalent binding caused the protein structure to unfold and the spatial conformation to become loose. Particle size and Zeta-potential measurements showed that the average particle size of conjugates(especially M-P-L-0.5 conjugate with PKP:LBG mass ratio of 1.0∶0.5) decreased and the surface charge increased, which could resist protein aggregation. The surface hydrophobicity and free thiol content of conjugates were significantly higher than those of physical mixtures (P<0.05), indicating that heating changed the tertiary structure of the protein and exposed hydrophobic groups, which was significantly related to the improvement of solubility and emulsifying properties of conjugates (M-P-L-0.5 increased by 0.41 times and 2.47 times, respectively, compared with PKP) (P<0.05). Pickering emulsions were prepared using conjugates as emulsifiers. The average particle size of the emulsion was positively correlated with the volume fraction of the oil phase. Microscopic structure observations showed that M-P-L-0.5 could adsorb on the surface of oil droplets to form an interface layer and stabilize the emulsion. The droplets of M-P-L-0.5 with a 60% oil phase were large and the gaps were small, and the emulsion stability was significantly higher than that of other emulsions (P<0.05). Moreover, this emulsion had good storage, acid-base, centrifugation, salt ion and temperature stability. This study clarified the mechanism by which LBG covalent modification improved the functional properties of PKP and successfully prepared highly stable Pickering emulsions, providing a basis for increasing the added value of pine kernel meal and expanding the application of PKP in food.

Abstract:The migration and invasion of tumor cells are important processes for the spread of tumors. In order to study the mechanism of consuming preserved eggs on the migration and invasion of human HepG2 cells, the effects of preserved eggs simulated gastrointestinal digests (PESD) on the migration and invasion of hepatoma HepG2 cells were studied by cell scratch test and Transwell cell invasion test. Western blot method was used to determine the expression of cell migration and invasion related factors in HepG2 cells induced by PESD. The process of epithelial-mesenchymal transition (EMT) induced by PESD was studied by real-time fluorescence quantitative PCR. The results showed that PESD inhibited the migration of HepG2 cells in a time-dose dependent manner. At 72h, the scratch healing rate of control group was 3 times higher than that of 4mg/mL PESD group. The invasion of HepG2 cells was inhibited in a dose-dependent manner. When PESD was 4mg/mL, the number of HepG2 cells passing through the membrane was only 20.33±3.06, which was significantly higher than that of the control group(P<0.001).Further study found that the inhibitory mechanism was that PESD affected the Wnt/ β-catenin signal pathway by reducing the expression of β-catenin protein. By increasing the expression of E-cadherin gene in HepG2 cells, reducing the expression of N-cadherin and Vimentin genes, inhibiting the process of EMT in HepG2 cells, increasing intercellular adhesion and the expression of matrix metalloproteinase and 9, the degradation of extracellular matrix and basement membrane was reduced, thus the migration and invasion of HepG2 cells was inhibitted. The research aimed to provide theoretical basis for exploring the health effects of preserved eggs.

Abstract:To investigate the effects of acetyl groups on the physicochemical properties of proanthocyanidins, the impact of Artemisia sphaerocephala Krasch glucomannan with different degrees of acetyl group substitution (DS) on the digestion characteristics of grape seed proanthocyanidins (GSP) was evaluated. Results indicated that the molecular weight of glucomannan remained stable during simulated digestion by using high-performance size-exclusion chromatography coupled with multi-angle laser light scattering. The results of content analyses of total phenol and total flavonoid and ultra-high performance liquid chromatography equipped with quadrupole time-of-flight tandem mass spectrometry, revealed that glucomannan enhanced the solubility of GSP in digestive liquids and increased the content of monomeric phenols such as catechin and epicatechin. Glucomannan significantly influenced the antioxidant and enzyme inhibitory activities of GSP in digestive liquids. Glucomannan improved DPPH radical scavenging activity of GSP, and the effect diminished as DS increased. Glucomannan enhanced inhibitory effects of GSP on α-glucosidase and lipase, with the impact strengthened as DS increased. Components of GSP, including catechin, epicatechin, gallocatechin, epigallocatechin gallate, catechin gallate, proanthocyanidins B1 and proanthocyanidins B2 and trimer, showed significant positive correlation with antioxidant and enzyme inhibitory activities. Among these, epicatechin gallate might play a key role. These findings suggested that acetylated glucomannan affected the digestion properties of GSP, with DS serving as a critical factor. An in-depth exploration of the interaction between proanthocyanidins and polysaccharides with specific structural characteristics could provide a theoretical basis for strategies to enhance the biological activities of proanthocyanidins.

Abstract:Jasmonic acid (JA) is one of the key pathways through which plants resist pathogenic fungi such as Botrytis cinerea, however, the changes and regulatory mechanisms of JA during the ripening of tomato fruit remain poorly understood. To explore this, the enzyme-linked immunosorbent assay was utilized to measure the JA content in the epidermal tissues of tomato fruits at various stages of maturity. Real-time fluorescence quantitative PCR was used to assess the expression levels of key JA biosynthesis genes, including 13-lipoxygenase (LOXD), 12-oxo-phytodienoate reductase 3 (OPR3), allene oxide synthase (AOS) and etc.. Virus-induced gene silencing technology was employed to elucidate the regulation of the basic helix-loop-helix protein encoding gene 47 (bHLH47) , a transcription factor linked to dual response to maturity and disease resistance, in JA synthesis of tomato fruits. The results indicated that the concentration of JA in green ripe tomato fruits was highest at 1.63nmol/L, whereas the concentration in red ripe fruits was lowest at 1.28nmol/L. In comparison to the small fruit stage, the expression of the OPR3 gene in red ripe fruits exhibited a significant downregulation, decreasing by 89.38%. This finding suggested that the synthesis of JA was influenced by the maturity of tomato fruits. In tomato fruits with bHLH47 gene silencing, JA concentration increased by 24.44%, the expression level of the OPR3 gene rose by 2.3 times compared to the control group, and the concentration of the 12-oxo-phytodiesterase (OPR) increased by 10.61%. The overall findings suggested that the transcription factor bHLH47 regulated JA synthesis by negatively influencing the expression of the OPR3 gene. This regulation subsequently impacted the resistance of tomato fruits to Botrytis cinerea during the ripening process. This study aimed to clarify the biological mechanisms that cause tomato fruits to become more susceptible to gray mold infection during ripening and offer theoretical insights for the management of post-harvest decay losses.

Abstract:Antimicrobial peptides are a type of peptide capable of exerting antibacterial functions by interacting with bacterial cell membranes or intracellular biomolecules, thereby disrupting bacterial physiological processes and ultimately leading to bacterial death. A novel deep learning model was constructed to screen antimicrobial peptides from soil metagenomic data and validated the screened peptides using techniques such as molecular docking and molecular dynamics simulations. The model demonstrated an outstanding performance with a precision of 98.7%, an accuracy of 96.5%, a recall rate of 91.9%, an F1-score of 95.2%, and a specificity of 99.2%, showcasing excellent efficiency, interpretability, and practical application value alongside robust generalization capabilities. After training, the model successfully identified several short peptides with significant antimicrobial potential, with a subset chosen for further investigation. The findings revealed that the screened peptide Gly-Thr-Ala-Trp-Arg-Trp-His-Tyr-Arg-Ala-Arg-Ser could effectively attach to the bacterial transcription regulator protein MrkH, exhibiting inhibitory effects on Klebsiella pneumoniae, Escherichia coli, and Staphylococcus aureus. This study aimed to provide a theoretical basis for the development and application of new antimicrobials in the food industry by integrating deep learning with molecular simulation technologies.

Abstract:To explore preparation method of citrus postharvest antifungal film, konjac glucomannan/curdlan-Bacillus sp. w176 fermentate (KCB) composite films were fabricated by combining natural polysaccharides with antifungal fermentate. The supernatant of w176 was subjected to vacuum freeze-drying to collect solid fermentate. The fermentate was combined with the polysaccharides to prepare composite solutions which was then cast and dried to form films. The w176 fermentate contained the Bacillus cells and its antifungal metabolites. The agar diffusion method for antimicrobial testing indicated that both the w176 fermentate and KCB composite films exhibited significant antifungal effects against Penicillium digitatum P44 and Penicillium italicum B3. The results of structural and property tests of the composite films showed that the surface and cross-sectional structure of the KCB composite films were continuous, uniform, and dense, with good compatibility between antifungal components and polysaccharides through non-covalent bonding. The w176 bacteria were uniformly distributed in the polysaccharide network. However, due to the hydrophilic nature of the antifungal components in the fermentate, the addition of w176 fermentate weakened the water resistance and mechanical properties of the composite films. The dissolution ratio (25.35%), water contact angle (71.7°), tensile strength (36.54MPa), elongation at break (8.86%), and water vapor and oxygen transmission rate (3.12kg·m-2·d-1 and 4.89g·m-2·h-1) of the KCB-5 composite film with a w176 fermentate concentration of 0.05g/L were significantly superior to other composite films. Comprehensive performance indicated its applicability for post-harvest preservation of citrus fruits. The antifungal experiment showed that KCB-5 composite film significantly inhibited the pathogenicity of pathogens B3 and P44 in postharvest preservation of citrus, which was of great significance to the citrus postharvest preservation industry.

Abstract:In light of the current situation that the grading of Lycium barbarum L. mainly relies on phenotypic characteristics such as color and fruit size, this study analyzed and evaluated the quality of dried Lycium barbarum L. based on the correlation analysis between functional components and phenotypic characteristics. A total of 224 samples of Lycium barbarum L. from different germplasms, harvesting batches, grades and origins were used to study the main functional components (total sugar, polysaccharide, betaine, total flavonoids and carotenoids), and phenotypic characteristics such as length, width, longitudinal area, and average single fruit weight and color indicators (L*, a* and b* values) of Lycium barbarum L.. Response of the main functional components and phenotype characteristics (fruit shape, size, color, etc.) of Lycium barbarum L. to different factors and the correlations between the quality indicators were analyzed. The total sugar content of Lycium barbarum L. was significantly and positively correlated with the fruit shape(P<0.05) and negatively correlated with the content of polysaccharides, betaine, total flavonoids and carotenoids. This means that Lycium barbarum L. with low total sugar content showed an overall trend of smaller fruit shape and higher contents of functional components. The color indexes L*, a* and b* were positively correlated with each other(P<0.05), L* and a* values were positively correlated with zeaxanthin dipalmitate and total carotenoids contents in different degrees. That was, the brighter and redder the Lycium barbarum L. fruit, the higher content of zeaxanthin dipalmitate and carotenoid. The carotenoids content of Lycium barbarum L. had a positive effect on the fruit brightness and redness. This study provided theoretical support for optimizing quality evaluation standards and breeding superior varieties of Lycium barbarum L..