Tropomyosin (TM), a major allergen in with strong immunobinding activity, underscores the importance of allergen-specific immunotherapy, for which hypoallergenic derivatives represent a promising strategy. In this study...Tropomyosin (TM), a major allergen in with strong immunobinding activity, underscores the importance of allergen-specific immunotherapy, for which hypoallergenic derivatives represent a promising strategy. In this study, the derivatives of TM from were prepared by selective deletion of B-cell epitopes and retention of T-cell epitopes and were named mTM. The hypoallergenicity of mTM was further confirmed in a mouse model of food allergy. Furthermore, the prophylactic efficiency of mTM was evaluated in a prophylactic mouse model for allergy. It was shown that mTM simultaneously attenuated IgE-mediated allergic responses, alleviated allergic responses via regulating Th1/Th2 balance, and enhanced the ratio of regulatory T cell-associated cytokines. Overall, the study provides a theoretical basis and research direction for the development of hypoallergenic derivatives for prophylactic administration in food allergies.
Cultured (CCS) improves sperm quality in patients with oligoasthenozoospermia (OAS), but its mechanisms remain unclear. A cyclophosphamide (CTX)-induced OAS mice model was employed to evaluate the effects of CCS on horm...Cultured (CCS) improves sperm quality in patients with oligoasthenozoospermia (OAS), but its mechanisms remain unclear. A cyclophosphamide (CTX)-induced OAS mice model was employed to evaluate the effects of CCS on hormone secretion, blood-testis barrier (BTB) integrity, testicular oxidative stress, and spermatogenic cell apoptosis. The steroidogenic factor-1 (SF-1)-mediated steroid hormone biosynthesis was further examined. Our results demonstrated that CCS elevated serum hormone levels, increased BTB-related protein expression, and inhibited spermatogenic cell apoptosis. Furthermore, CCS upregulated the protein expression of steroidogenic enzymes and SF-1 in testes. In vitro, CCS extract enhanced testosterone production and steroidogenic enzyme expression in CTX-injured TM3 cells, concomitant with the upregulation of SF-1. Moreover, SF-1 knockdown reversed the effect of CCS extract on testosterone production. In conclusion, CCS improves OAS by enhancing testosterone production primarily via activation of SF-1-mediated steroid hormone biosynthesis. The increased testosterone contributes to maintaining BTB integrity and inhibiting testicular cell apoptosis.
β-Galactosidases are widely used to produce lactose-free dairy products, but efficient lactose hydrolysis under refrigerated conditions remains limited by the low activity of many commercial enzymes. In this study, a col...β-Galactosidases are widely used to produce lactose-free dairy products, but efficient lactose hydrolysis under refrigerated conditions remains limited by the low activity of many commercial enzymes. In this study, a cold-adapted β-galactosidase OUC-JST from was characterized and engineered. OUC-JST exhibited maximal activity at 20 °C and pH 7.0. Structure-guided mutagenesis identified R110I, which shifted the optimal temperature to 15 °C, while K1023D increased activity approximately 3-fold. The combined variant R110I/K1023D (OUC-JST-N) retained the optimal temperature of 15 °C and exhibited 2.80-fold higher specific activity than the wild type. Molecular dynamics simulations suggested that OUC-JST-N may modulate correlated motions and active-site flexibility, providing a possible explanation for its improved performance. In pasteurized whole milk, OUC-JST-N hydrolyzed 98.37% of lactose at 10 °C within 48 h, leaving 812.25 mg/L residual lactose. This study highlights the potential of the cold-adapted β-galactosidase OUC-JST and its variant in the dairy processing industry as a solution to lactose intolerance.
In recent years, nutrition and health have become topics of widespread societal concern, and galactooligosaccharides (GOS) have garnered significant interest as prebiotics. Composed of one to nine galactose units termina...In recent years, nutrition and health have become topics of widespread societal concern, and galactooligosaccharides (GOS) have garnered significant interest as prebiotics. Composed of one to nine galactose units terminated by a glucose molecule, GOS exhibit remarkable thermostability, high water solubility, strong humectancy, and low caloric value. They provide various physiological benefits, including prebiotic activity, gut health maintenance, antibacterial, antiadhesive, and anti-inflammatory effects, as well as systemic regulation of blood sugar, lipids, immune homeostasis, and metabolic diseases. These properties support broad applications in the food industry, breeding field, material science, and medical science. Enzymatic production using β-galactosidases is the primary industrial method for GOS synthesis. This review provides an overview of recent advances in β-galactosidase biochemistry, crystal structures, catalytic mechanisms, molecular modification, immobilization, and heterologous expression, along with GOS physiological functions, production, applications, and whey utilization. Future opportunities for GOS research and development are also briefly discussed.
Wang D, Wu B, Liu X
… +3 more, Yu Y, Kang Q, Bai L
J Agric Food Chem
· 2026 Jul · PMID 42333452
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Transglutaminase (TGase) from is widely used in food processing to improve the texture properties of food products, but its broader application is constrained by low titers. Herein, through a quantitative analysis of th...Transglutaminase (TGase) from is widely used in food processing to improve the texture properties of food products, but its broader application is constrained by low titers. Herein, through a quantitative analysis of the intracellular amino acid pool, we identified an insufficient supply of several amino acids as the primary bottleneck for TGase overproduction. To address this limitation, we overexpressed the biosynthetic pathways of seven critically insufficient amino acids, which increased the TGase titer by 46.3% (from 41.0 to 60.0 U/mL). Additionally, repressing the expression of nonessential high-abundance proteins reduced the metabolic competition for amino acids, boosting the TGase titer by 57.5% (from 40.0 to 63.0 U/mL). Combining both strategies yielded a maximum titer of 66.9 U/mL, which is a 62.4% improvement over the starting strain. This study provides a foundation for efficient TGase production and highlights precursor flux manipulation as a general strategy for enzyme overproduction.
Mercury sulfide nanoparticles (HgS) are widespread yet poorly understood Hg species in contaminated areas. Here, we present a 2.5-year simulated experiment tracking the persistence, transformation, and bioavailability of...Mercury sulfide nanoparticles (HgS) are widespread yet poorly understood Hg species in contaminated areas. Here, we present a 2.5-year simulated experiment tracking the persistence, transformation, and bioavailability of HgS relative to larger HgS forms in soil-rice systems. We demonstrate that HgS persist in soils with minimal physicochemical change but exhibit high methylation potential and rice uptake. Soil amended with HgS yielded 13- to 26-fold higher methylmercury (MeHg) concentrations than bulk HgS treatments, resulting in 14- to 92-fold and 5.6- to 68-fold increases in MeHg and inorganic Hg in brown rice, respectively. Mechanistically, this sustained bioavailability arises from the synergistic effects of HgS translocation within rice tissues, reactive Hg release, and enhanced methylation. Our findings identify HgS as a persistent and highly bioavailable Hg pool in paddies, fundamentally driving long-term Hg transformation and accumulation in rice. This work redefines Hg biogeochemical cycling in agricultural ecosystems and underscores the critical need to incorporate HgS into food safety assessments.
Plants emit volatile organic compounds (VOCs) under stress, generating complex blends that mediate defense and insect host location. β-Ocimene is a widespread stress-associated monoterpene induced by herbivory, mechanica...Plants emit volatile organic compounds (VOCs) under stress, generating complex blends that mediate defense and insect host location. β-Ocimene is a widespread stress-associated monoterpene induced by herbivory, mechanical damage, heat, and drought, and influences insect behavior; however, its molecular detection as a kairomone remains unclear. Using the Eurasian longhorn beetle , we found that females are significantly attracted to β-ocimene. Antennal transcriptomics and expression profiling identified candidate odorant-binding proteins (OBPs), among which MsutOBP13 showed high binding affinity for β-ocimene. Structural modeling, molecular docking, and mutagenesis identified Phe49 as a key residue for ligand recognition. RNA interference further confirmed that MsutOBP13 contributes to β-ocimene-mediated behavioral responses. These results reveal a molecular basis for β-ocimene detection and identify this volatile as a host-location cue in a wood-boring herbivore.
Regional diversity may influence the composition of human milk fat globule membrane (HMFGM) protein composition and protein glycosylation. Here, we first systematically evaluated the efficiency and stability of six metho...Regional diversity may influence the composition of human milk fat globule membrane (HMFGM) protein composition and protein glycosylation. Here, we first systematically evaluated the efficiency and stability of six methods for the extraction of HMFGM proteins from breast milk. The optimal method (methanol-chloroform precipitation of MFGM proteins, followed by solubilization in 0.4% SDS lysis buffer) was further applied to the proteome and N-glycoproteome analysis of 49 breast milk samples from five regions across China. Proteomic analysis revealed the geographically divergent expression of BST2, COCH, FUCA1 and FGFBP1 in MFGM. The parallel N-glycoproteomic profiling identified 4914 site-specific N-glycans mapping to 689 glycoproteins, 699 glycosylation sites, and 420 glycan structures. This multiomics study reveals region-specific variations in the molecular composition of HMFGM proteins and their N-glycoprotein derivatives, elucidating the structural and functional dynamics of the human HMFGM in representative regions of China.
Microplastic-derived dissolved organic matter poses an emerging threat to terrestrial ecosystems. Here, we investigated the effects of PP-DOM and PS-DOM (at concentrations of 0.1, 1, and 10 mg/L) on Chinese cabbage (), p...Microplastic-derived dissolved organic matter poses an emerging threat to terrestrial ecosystems. Here, we investigated the effects of PP-DOM and PS-DOM (at concentrations of 0.1, 1, and 10 mg/L) on Chinese cabbage (), planted soils, and soil microbial communities. PP-DOM at 1 mg/L increased soil TOC, TN, and phosphorus, enriched opportunistic pathogens (), and affected iron outer membrane receptor proteins, while 1 mg/L PS-DOM enhanced available nitrogen and nitrogen-transforming bacteria (, , and ). Both of them increased the abundance of three carbon metabolism-related genes (, , ). PP-DOM stimulated plant growth at 0.1 mg/L but inhibited growth at higher concentrations; PS-DOM exhibited the opposite trend. PP-DOM at higher concentrations reduced chlorophyll and suppressed CAT/superoxide dismutase activities, while high-level PS-DOM increased chlorophyll and carotenoids with weaker oxidative damage. Transcriptomic analysis revealed that MP-DOM modulated the plant photosynthetic and amino acid metabolism pathways. Collectively, MP-DOM chemical composition critically shapes soil-microbe-plant ecological outcomes.
Cell cryopreservation is essential for food biotechnology and cell-based bioprocessing. However, the widely used cryoprotectant dimethyl sulfoxide (DMSO) shows dose-dependent cytotoxicity, motivating safer alternatives....Cell cryopreservation is essential for food biotechnology and cell-based bioprocessing. However, the widely used cryoprotectant dimethyl sulfoxide (DMSO) shows dose-dependent cytotoxicity, motivating safer alternatives. Here, food-derived antifreeze peptides isolated from skin (TsAFP) were evaluated as primary cryoprotectants using stem cells as a representative cellular model. At 15 mg/mL, TsAFP achieved a 70% replacement of DMSO while maintaining post-thaw cell growth, metabolic activity, adhesion, and multilineage differentiation. TsAFP increased solution viscosity and reduced water activity, effectively inhibiting ice growth and recrystallization. It also adsorbed onto cell membranes to form a protective interfacial layer. Molecular dynamics simulations revealed that TsAFP disrupted water ordering through stable hydrogen-bond interactions. Together, TsAFP provides extracellular and interfacial protection, while low-dose DMSO contributes intracellular protection, resulting in coordinated cryoprotection. TsAFP offers a safer, food-derived strategy to reduce DMSO reliance.
The unique characteristics of nanobodies make them ideal candidates for diagnostic and therapeutic applications. Introducing additional disulfide bonds is one of the most effective ways to enhance a nanobody's stability....The unique characteristics of nanobodies make them ideal candidates for diagnostic and therapeutic applications. Introducing additional disulfide bonds is one of the most effective ways to enhance a nanobody's stability. However, it always negatively affects the nanobody's affinity and expression in host cells. In this study, an anticarbaryl nanobody (NbCBR) was used as a model to investigate the strategy of additional disulfide bond introduction. The crystal structure of the NbCBR-carbaryl complex was determined, based on which molecular dynamics simulations and virtual screening were carried out. Seven disulfide bond-introduced mutants were then constructed. Compared with the wild-type nanobody, all the mutants retained good sensitivity. The results demonstrate that introducing disulfide bonds between two β-sheets or anchoring the flexible CDR3 to a β-sheet can significantly enhance the nanobody's stability under thermal, organic solvent, and pH stress conditions. This study provides a theoretical basis for rational engineering of nanobodies to improve their stability, potentially expanding their further applications.
Food allergy (FA), defined as abnormal immunological response to certain food proteins, is a serious issue in food safety and public health. Dendritic cells (DCs) orchestrate tolerance or allergic reactions to food prote...Food allergy (FA), defined as abnormal immunological response to certain food proteins, is a serious issue in food safety and public health. Dendritic cells (DCs) orchestrate tolerance or allergic reactions to food proteins. Accumulating researches show that dietary components can intervene in the progression of FA by modulating DCs function. However, existing reviews have largely focused on effector cells, such as T cells and mast cells, and the key role of DCs in how dietary components influence FA is still lacking. Therefore, the aim of the review is to discuss the antiallergic mechanism based on dietary components-DCs-FA axis. It outlines the functions of DCs in FA and elucidates how specific dietary components, including vitamins, fatty acids, polyphenols, dietary fiber, and probiotics, influence the development of FA by regulating DCs. This review offers novel insights into DCs-mediated mechanisms and facilitates the development of targeted dietary intervention for FA.
Piperidine alkaloids in venom are natural substances with insecticidal activity. Their structural diversity provides a model for investigating structure-activity relationships to develop novel targeted insecticides. Usi...Piperidine alkaloids in venom are natural substances with insecticidal activity. Their structural diversity provides a model for investigating structure-activity relationships to develop novel targeted insecticides. Using the agricultural pest , this study systematically investigated the differential effects of piperidine alkaloid / isomerization and side-chain elongation on toxicity and mode of action. Results revealed that -configuration alkaloids exhibited significantly higher toxicity than their -configuration counterparts. Side-chain elongation enhanced the toxicity of -configuration alkaloids (C11-C13) but did not affect their -configuration. Through RNAi and CRISPR/Cas9, we found that -configuration alkaloids target the α2 and β1 nAChR subunits, while the -configuration alkaloids target the α1, α2, and β1 nAChR subunits. Furthermore, α1, α2, and β1 subunits also participated in the resistance mechanism of to imidacloprid. Combined with predictions of alkaloid-target binding modes, our findings provide crucial mechanistic insights for optimizing piperidine alkaloid structures and designing specifically targeted insecticides.
Aphids are important worldwide pests in crop production. Steroidal Piperidone Compound exhibits high insecticidal activity against aphids, but its aphidicidal mechanism remains unclear. In this study, drug affinity resp...Aphids are important worldwide pests in crop production. Steroidal Piperidone Compound exhibits high insecticidal activity against aphids, but its aphidicidal mechanism remains unclear. In this study, drug affinity responsive target stability (DARTS) was used to identify pyruvate kinase (PK) as the target protein of in . qRT-PCR and enzyme activity assays showed that upregulated PK expression and enhanced its enzymatic activity. RNA interference (RNAi)-mediated silencing of PK significantly reduced the insecticidal toxicity of , whereas PK complementation restored this toxicity. Molecular docking and site-directed mutagenesis verified Ser142 as the key binding site, and enzymatic kinetic analysis confirmed that acts as a noncompetitive activator of PK. This study clarifies the molecular mechanism whereby exerts its insecticidal activity by noncompetitively activating PK, supporting PK as a valuable target for novel insecticide design.
Sugarcane smut, caused by , is a globally destructive disease that severely threatens sugarcane production. In this study, we isolated an endophytic bacterium, ZC2-4, which exhibits inhibitory activity against the mycel...Sugarcane smut, caused by , is a globally destructive disease that severely threatens sugarcane production. In this study, we isolated an endophytic bacterium, ZC2-4, which exhibits inhibitory activity against the mycelial and haploid growth of . Whole-genome sequencing and bioinformatics analysis predicted 13 secondary metabolite biosynthetic gene clusters, and the antifungal compound was subsequently identified as Iturin A-4 through high-performance liquid chromatography and nuclear magnetic resonance spectroscopy. Transcriptomic analysis and physiological assays revealed antagonistic mechanisms of Iturin A-4 against haploid growth including induction of oxidative stress and interference with cell-cycle progression. Scanning and transmission electron microscopy observations further demonstrated that Iturin A-4 inhibits teliospore germination. Remarkably, Iturin A-4 exhibits broad-spectrum antifungal activity against various fungal pathogens. Collectively, this study identifies ZC2-4 as a promising biocontrol agent and elucidates the antifungal mechanisms of Iturin A-4, providing a foundation for sustainable management of sugarcane smut.
Dysphagia is a common clinical symptom, often accompanied by insufficient lubrication of the oral cavity and pharynx, which reduces eating efficiency and nutrient intake, increases the risk of complications such as aspir...Dysphagia is a common clinical symptom, often accompanied by insufficient lubrication of the oral cavity and pharynx, which reduces eating efficiency and nutrient intake, increases the risk of complications such as aspiration pneumonia. Artificial saliva is a product designed to mimic natural saliva, commonly used to provide oral lubrication. However, few studies have demonstrated its efficacy in improving dysphagia. Epidemiological data clarify the characteristics of populations for dysphagia, while the process of swallowing dysfunction induced by etiological factors provides a clinical basis for the application of artificial saliva. This review highlights the importance of artificial saliva in the management of dysphagia, summarizes the current status of research on artificial saliva in recent years, and elucidates the mechanisms underlying the lubrication behavior of artificial saliva from the perspective of the salivary multilayer through its interaction with the salivary conditioning film (SCF), thereby paving the way for next generation artificial saliva.
Acid phosphatases primarily catalyze phosphate ester hydrolysis, yet some members exhibit promiscuous phosphotransferase activity, enabling substrate phosphorylation using inexpensive inorganic phosphate. This catalytic...Acid phosphatases primarily catalyze phosphate ester hydrolysis, yet some members exhibit promiscuous phosphotransferase activity, enabling substrate phosphorylation using inexpensive inorganic phosphate. This catalytic promiscuity provides attractive biocatalysts for the sustainable production of phosphorylated food-related compounds, but substrate-dependent activity variation makes efficient enzyme identification and engineering challenging. Here, we developed TSCF, a strategy that integrates machine learning with transition-state conformational features to mine enzyme sequences and guide sequence optimization using transition-state models. For inosine 5'-phosphate formation, TSCF showed strong predictive capability, with 70.0% of 20 designed mutants exhibiting 1.2-6.4-fold higher activity than their wild-type enzymes. Incorporation of molecular dynamics simulations further increased screening accuracy to 77.8%. Application to L-ascorbic acid phosphorylation identified effective mutants with 75.0% accuracy and 71.1% conversion, demonstrating the generality of TSCF for improving acid phosphatase-catalyzed phosphotransfer.
Silva E, Santana Aguiar MC, Vilhena Araújo E
… +5 more, França P, Berlinck RGS, Ballester AR, González-Candelas L, Fill T
J Agric Food Chem
· 2026 Jul · PMID 42322288
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, the causal agent of citrus green mold, remains the most destructive postharvest pathogen of citrus worldwide. Recent advances have greatly expanded our understanding of the molecular dialogue between and citrus hosts,..., the causal agent of citrus green mold, remains the most destructive postharvest pathogen of citrus worldwide. Recent advances have greatly expanded our understanding of the molecular dialogue between and citrus hosts, revealing coordinated virulence strategies involving cell wall-degrading enzymes, major facilitator superfamily transporters, transcription factors, and secondary metabolism, alongside host defenses mediated by phytohormones and specialized metabolites. This review integrates genomic, transcriptomic, metabolomic, and functional genetic discoveries, including CRISPR/Cas9 and -mediated transformation, which have accelerated the characterization of fungal pathogenicity and host resistance. We further assess biological control as a sustainable alternative to chemical fungicides, emphasizing complementary mechanisms such as niche competition, antibiosis, volatile organic compound (VOC) production, biofilm formation, iron sequestration, lipopeptide synthesis, and induction of host defenses. In addition, we highlight microbiome-informed strategies and the design of synthetic microbial communities (SynComs) as promising next-generation approaches to enhance efficacy, stability, and ecological resilience in citrus postharvest disease management.
The development of anticoagulants targeting coagulation factor XIa (FXIa) is a promising strategy to reduce thrombotic risk while minimizing bleeding complications. However, FXIa inhibitory peptides derived from the deep...The development of anticoagulants targeting coagulation factor XIa (FXIa) is a promising strategy to reduce thrombotic risk while minimizing bleeding complications. However, FXIa inhibitory peptides derived from the deep sea remain largely unexplored. In this study, pepsin hydrolysates from the deep-sea mussel exhibited anticoagulant effects through prolongation of activated partial thromboplastin time (APTT) and reducing erythrocyte and platelet aggregation in a zebrafish thrombosis model. Through bioactivity-guided fractionation combined with AI-assisted virtual screening, three novel FXIa inhibitory peptides were identified: VVTRVVR (IC = 523 μM), TVTRLSD (IC = 683 μM), and VSQPRSTL (IC = 695 μM). These peptides selectively inhibited FXIa without affecting FXa or thrombin and prolonged APTT. Structural analyses using AlphaFold3 modeling, molecular dynamics simulations, and quantum chemical calculations revealed that binding stability was driven by electrostatic interactions with Asp189 in the S1 pocket. This work provides a strategy for discovering anticoagulant peptides.
Liver diseases are a major global health burden with an occult onset and atypical symptoms, hindering early diagnosis. Herbal medicines feature low toxicity and multitarget effects; oxymatrine (OMT) from possesses anti-...Liver diseases are a major global health burden with an occult onset and atypical symptoms, hindering early diagnosis. Herbal medicines feature low toxicity and multitarget effects; oxymatrine (OMT) from possesses anti-inflammatory, antioxidant, antiapoptotic, and antitumor activities, with prominent hepatoprotective effects. This systematic review explores the impact and molecular mechanisms of oxymatrine in managing various liver ailments, including liver injury, nonalcoholic fatty liver disease (NAFLD), liver fibrosis, and hepatocellular carcinoma (HCC), based on the findings from in vitro and in vivo studies. Using scientific databases such as PubMed, Web of Science, and Science Direct, studies were analyzed focusing on oxymatrine's pharmacological actions in liver disease. The search incorporated terms including "oxymatrine," "hepatoprotection," and "liver disease." Oxymatrine mediates liver protection by modulating oxidative stress, suppressing inflammation, promoting lipid metabolism, and inhibiting fibrosis. Additionally, it exhibits tumor-suppressive effects through apoptosis induction and pathway modulation.