Soybean productivity is significantly impacted by drought and salt stress, yet the precise molecular mechanisms responsible for abiotic stress tolerance mediated by the SUMO protease GmOTSa remain unclear. To address thi...Soybean productivity is significantly impacted by drought and salt stress, yet the precise molecular mechanisms responsible for abiotic stress tolerance mediated by the SUMO protease GmOTSa remain unclear. To address this gap, -overexpressing and CRISPR/Cas9-edited knockout soybean lines were developed to investigate its function and regulatory pathways. boosts drought and salt tolerance by scavenging reactive oxygen species, modulating ABA-mediated stomatal movement, and increasing the expression of stress-responsive genes. Analysis using label-free quantitative proteomics revealed an enrichment of pathways related to linoleic acid metabolism under the stress conditions. Furthermore, GmOTSa physically interacts with the 13-lipoxygenase GmLOX31, facilitating its deSUMOylation, which, in turn, stabilizes the GmLOX31 protein and enhances jasmonic acid biosynthesis. Co-overexpression of and results in a synergistic improvement in stress tolerance. This investigation unveils a novel regulatory module involving GmOTSa and GmLOX31 deSUMOylation, connecting deSUMOylation to lipid metabolism and the adaptation of soybeans to abiotic stress.
Bamboo sugars such as glucose and cellobiose can be transformed into valuable biochemicals. Cellobiose, a glycosyl precursor, enables the synthesis of C-glycosyl flavonoids, such as vitexin, a bioactive compound in bambo...Bamboo sugars such as glucose and cellobiose can be transformed into valuable biochemicals. Cellobiose, a glycosyl precursor, enables the synthesis of C-glycosyl flavonoids, such as vitexin, a bioactive compound in bamboo extracts. This study engineered a recombinant strain to produce vitexin from apigenin and cellobiose. Optimizing conditions increased the vitexin production to 3092 mg/L. To reduce the production cost of vitexin, the enzymatic hydrolysate of alkali-pretreated bamboo was utilized to replace cellobiose, serving as both the glycosyl donor precursor and the carbon source for the recombinant strain to produce vitexin. By optimization of the conditions of bamboo pretreatment and enzymatic hydrolysis with peptone, 100 g of bamboo can produce 18.1 g of cellobiose and 11.3 g of glucose, and the cellobiose and glucose can be used to produce 37.9 g of vitexin in the recombinant strain. This study provides a novel method for the production of vitexin using enzymatic hydrolysis of bamboo cellulose.
The double mutant MsE of the methyltransferase EgtD from can catalyze the N-terminal trimethylation of L-tryptophan to produce the medicinal indole alkaloid L-hypaphorine. Structural comparison, loop deletion, and alani...The double mutant MsE of the methyltransferase EgtD from can catalyze the N-terminal trimethylation of L-tryptophan to produce the medicinal indole alkaloid L-hypaphorine. Structural comparison, loop deletion, and alanine scanning revealed that Loop34-39 and Loop163-168 are critical for MsE activity and substrate binding. Saturation and iterative saturation mutagenesis targeting hotspot residues in these loops yielded mutant M5 (P35C/K36R/F38L/T163G/T168F). Compared with the original enzyme MsE, M5 exhibits a 7.11-fold increase in relative activity and achieves a conversion rate of 58.93%. Molecular dynamics simulations, CAVER tunnel analysis, and ligand dissociation pathway studies reveal that M5 modulates the hydrogen bond network of the active site and remodels the conformation of the substrate access tunnel, thereby transforming the substrate tunnel into a methylation reaction microcompartment with a "capture-release" function. This study provides a theoretical and practical basis for loop optimization engineering and enzyme functional modification.
Engineering chitinases with simultaneous improvements in catalytic efficiency and thermostability remains challenging. Here, we present a computational strategy targeting the insertion domain (CID), a noncatalytic auxili...Engineering chitinases with simultaneous improvements in catalytic efficiency and thermostability remains challenging. Here, we present a computational strategy targeting the insertion domain (CID), a noncatalytic auxiliary region. Using Chi18A_ΔChBDs as the starting scaffold, folding free energy calculations identified a stabilizing mutation, A347P, within the CID hydrophobic core, which significantly enhanced thermostability by extending the half-life at 45 °C by 56 min. Based on this stabilized scaffold, evolutionary coupling analysis and stability predictions guided the design of secondary mutations, yielding several double mutants with improved catalytic performance. The best variants (A347P/A137S, A347P/A158G, and A347P/E219A) exhibited increases in specific activity of up to 99% compared with the wild-type and substantially prolonged half-lives at 50 °C. Notably, A347P/A137S produced 4.9-fold more GlcNAc during colloidal chitin hydrolysis. These results demonstrate that CID-targeted engineering provides an effective strategy for optimizing multidomain chitinases.
Sansalvamides are a class of bioactive cyclodepsipeptides recognized for their potent antitumor properties. In this study, genome mining of an endophytic fungus sp. R1 led to the identification of a unique nonribosomal...Sansalvamides are a class of bioactive cyclodepsipeptides recognized for their potent antitumor properties. In this study, genome mining of an endophytic fungus sp. R1 led to the identification of a unique nonribosomal peptide synthetase (NRPS) gene cluster (), which consists of 5 core genes and 14 auxiliary tailoring genes. Chemical investigation of strain R1 resulted in discovery of one new noncyclic sansalvamide derivative together with three analogues (-). Using CRISPR-Cas9-mediated gene knockout, we disrupted the cluster for the first time, abolishing production of compounds - and confirming its functional indispensability. Transcriptomic analysis unveiled a GDSL-like lipase/acylhydrolase () responsible for catalysis of a key ring-opening step followed by the production of and . Antifungal assays revealed that compounds and possessed pronounced activity against the phytopathogenic fungus with inhibition rates of 66.7 and 77.0%, respectively. In addition, compound exhibits excellent inhibitory activity against tumor cell lines HCT-116 and MCF-7/ADR with IC values of 0.71 and 12.50 μM, respectively, indicating its potential application in agrochemical and pharmaceutical industry. Simultaneously, these results showed that the role of the macrocyclic scaffold of sansalvamides is essential for antifungal effect and the -methylation of these chemicals could improve efficiency.
The pathogen causes postharvest sour rot disease in citrus fruit worldwide, severely impacting the commercial development and economic value of citrus. However, research on commenced relatively late, and the pathogenic...The pathogen causes postharvest sour rot disease in citrus fruit worldwide, severely impacting the commercial development and economic value of citrus. However, research on commenced relatively late, and the pathogenic mechanisms are incompletely understood. This study successfully constructed # and # overexpressing lines via PEG-mediated protoplast transformation. Compared to WT, the overexpressing strains enhanced pathogenicity by increasing chitin levels through the elevated expression of , , and . They promoted degradation of citrus protopectin, enhanced expression of key membrane lipid metabolism genes and , and increased malondialdehyde (MDA) accumulation, thereby compromising cell membrane integrity. This demonstrated that and represent critical targets for driving the progression of sour rot, filling gaps in our understanding of the pathogenic mechanisms of . These findings hold significant implications for future drug target development and the advancement of integrated management strategies against sour rot.
Drought stress inhibits the growth and development of , and WUSCHEL-related homeobox (WOX) transcription factors participate in plant growth and abiotic stress responses. However, the mechanism of the WOX-regulated droug...Drought stress inhibits the growth and development of , and WUSCHEL-related homeobox (WOX) transcription factors participate in plant growth and abiotic stress responses. However, the mechanism of the WOX-regulated drought response in remains to be elucidated. Here, we identified the drought-responsive WOX transcription factor ArWOX11. Overexpression of weakened drought resistance in and . ArWOX11 directly binds to the promoter (protein phosphatase 2C) and transcriptionally represses its expression. ArWOX11 interacts with itself and a novel regulator at the plasma membrane (NRPM) protein, ArNRPM3, in plants. Meanwhile, coexpression of ArWOX11 and ArNRPM3 enhanced the water loss rate and suppressed expression more than ArWOX11 alone, while abscisic acid (ABA) alleviated their inhibitory effect on . Overall, this study elucidates the function of the ArWOX11-ArNRPM3 module in the drought response, providing insights into the action mechanism of WOX in plant drought adaptation.
The evolution of volatile esters is a key determinant of the sensory profile of young white wines during the storage. This study evaluates the stability of 17 esters in 32 commercial wines under forced aging conditions....The evolution of volatile esters is a key determinant of the sensory profile of young white wines during the storage. This study evaluates the stability of 17 esters in 32 commercial wines under forced aging conditions. Although ester hydrolysis is typically associated with initial ester concentrations and pH conditions during storage, the results show that the wine matrix plays a major role in modulating reaction rates. Multivariate analysis identified metals, especially Fe and Mn, as significant drivers of ester degradation, suggesting a catalytic effect. In contrast, K and Mg showed an influence comparable to that of the pH. Overall metal composition and pH proved to be better predictors of shelf life than initial ester concentrations. These findings challenge traditional assumptions about ester equilibrium and highlight the importance of managing the metal content to preserve freshness and fruity aromas in young white wines over time.
The scarab beetle is a significant agricultural pest, causing crop damage through adult feeding and larval root destruction. Targeting its sex pheromone system provides a promising eco-friendly pest control strategy. Ph...The scarab beetle is a significant agricultural pest, causing crop damage through adult feeding and larval root destruction. Targeting its sex pheromone system provides a promising eco-friendly pest control strategy. Pheromone-binding proteins (PBPs) are essential in the process of pheromone detection. We functionally characterized AcorOBP4 and AcorOBP15, which are predominantly expressed in male antennae. AcorOBP15 exhibited the highest binding affinity toward the sex pheromone ()-japonilure, while showing comparatively weaker binding to host plant volatiles, suggesting a pheromone-dominant role in odorant detection. RNAi knockdown of significantly reduced the electrophysiological and behavioral responses of male beetles to ()-japonilure. Molecular docking revealed that AcorOBP15 interacts with ()-japonilure through a combination of hydrophobic interactions and a single hydrogen bond. Finally, mutagenesis of the hydrogen-bonding site Ser121 markedly reduced pheromone binding affinity. These findings highlight AcorOBP15 as a pheromone-dominant, multifunctional PBP that may contribute to the coordinated perception of mating and host-related cues at the peripheral olfactory level.
Freshwater fish is widely consumed, yet raw or undercooked freshwater fish can transmit foodborne zoonotic pathogens. Clonorchiasis, caused by , is a major foodborne parasitic disease associated with hepatobiliary injury...Freshwater fish is widely consumed, yet raw or undercooked freshwater fish can transmit foodborne zoonotic pathogens. Clonorchiasis, caused by , is a major foodborne parasitic disease associated with hepatobiliary injury and fibrosis. The biological functions and mechanisms underlying liver injury mediated by -derived extracellular vesicles (EVs) encapsulated microRNAs in fibrogenic remodeling remain poorly defined. Here, we isolated and characterized EVs from adult , confirmed their uptake by hepatic stellate cells (HSCs), and identified as a EV-enriched microRNA. enhances HSC activation and upregulates fibrogenic markers, including α-SMA, COL1A1, and COL3A1, both and in a mice infection model. Bioinformatic prediction and dual-luciferase assays validated TRIM35 as a target of . TRIM35 silencing phenocopied overexpression and increased PI3K/AKT phosphorylation, whereas inhibition restored TRIM35 and attenuated fibrogenic markers. Collectively, EV-associated promotes HSC activation via the TRIM35-PI3K/AKT axis, suggesting a potential target for antifibrotic intervention in clonorchiasis. These findings provide mechanistic insight into how infection acquired from raw or undercooked freshwater fish harboring infective metacercariae can lead to hepatobiliary injury and fibrogenic remodeling, supporting food safety risk assessment and future prevention of fish-borne parasitic hazards.
ζ-Carotene is a key biosynthetic intermediate of lycopene that is widely distributed in plants and fungi but difficult to purify, which has limited detailed physicochemical and biological studies. Here, we established a...ζ-Carotene is a key biosynthetic intermediate of lycopene that is widely distributed in plants and fungi but difficult to purify, which has limited detailed physicochemical and biological studies. Here, we established a purification strategy that affords four ζ-carotene geometric isomers (all--, 9, 9,9', and 9,13,9'-isomers) in high purity and elucidated their structures fully using nuclear magnetic resonance spectroscopy. Photochemical experiments showed that, unlike many carotenoids that photoisomerize toward the all--form, ζ-carotene yields a photostationary mixture dominated by the 9-isomer. For the major -isomers, (9)- and (9,9')-ζ-carotene, we determined solvent-dependent spectral properties, color parameters, and ultraviolet (UV)-shielding capacities, revealing remarkably strong UV-A shielding. These -isomers also exhibited potent singlet oxygen quenching and pronounced inhibition of fluorescent advanced glycation end-product formation, together with detectable hydroxyl radical scavenging. Our findings provide fundamental data supporting ζ-carotene as a promising natural UV-A filter and skin health-promoting functional ingredient.
Kiwifruit presents a growing global health concern, characterized by IgE-mediated type I hypersensitivity reactions ranging from mild oral symptoms to severe anaphylaxis. This allergy is driven by five major allergenic p...Kiwifruit presents a growing global health concern, characterized by IgE-mediated type I hypersensitivity reactions ranging from mild oral symptoms to severe anaphylaxis. This allergy is driven by five major allergenic proteins (Act d 1, 2, 5, 8, and 12), which exhibit significant cross-reactivity, contributing to conditions such as latex-fruit syndrome and pollen-food allergy syndrome. The structural heterogeneity and epitope variations among kiwifruit allergens directly influence the diversity and severity of clinical reactions. Accurate epitope mapping may improve diagnostic accuracy, clarifies cross-reactivity mechanisms and assist in risk assessment, while food processing techniques modify allergenic protein structures. Understanding how processing affects epitope accessibility is essential for developing low-allergenic kiwifruit products and improving food safety. This review synthesizes current understanding of kiwifruit allergy, emphasizing the structural and immunological basis of its major allergens, and assesses how food processing can mitigate allergenicity to guide clinical management and product innovation.
Stilbenoids are metabolites with inhibitory activity against plant pathogens, such as , the causal agent of grapevine downy mildew. However, their activity is strongly reduced under UV-A exposure because of photodegradat...Stilbenoids are metabolites with inhibitory activity against plant pathogens, such as , the causal agent of grapevine downy mildew. However, their activity is strongly reduced under UV-A exposure because of photodegradation. This study explores strategies to improve stilbenoid photostability in a grapevine cane extract (CE) while preserving its bioactivity. Among these, encapsulating CE stilbenoids in a cellulose nanocrystal-stabilized oil-in-water emulsion achieved 53% encapsulation efficiency and proved most effective, limiting stilbenoid photodegradation to 37.5% after 1 h of UV-A exposure, compared to 73.9% in the control. However, in this case, the biological activity against downy mildew was loosened. Combinations of CE with a flavonoid (hesperidin methyl chalcone or rutin) also improved stilbenoid photostability, although less markedly, while still significantly preserving the activity of CE against . These findings highlight that such photoprotection strategies support the realistic use of stilbenoid extracts as eco-friendly alternatives for sustainable disease management in grapes.
Conventional ACE-inhibitory peptide production methods suffer from low yield, suboptimal activity, and extensive optimization. This study introduces DeepAngio, a novel model to address these limitations and enable target...Conventional ACE-inhibitory peptide production methods suffer from low yield, suboptimal activity, and extensive optimization. This study introduces DeepAngio, a novel model to address these limitations and enable targeted release. DeepAngio demonstrates high performance in screening (98.80% accuracy, 99.63% precision, and 96.44% specificity) and classification (88.24% accuracy, 86.76% precision, and 95.16% specificity) validated through 10-fold cross-validation. Moreover, DeepAngio facilitated the development of a method for directional preparation of tuna meat hydrolysates with notable ACE inhibitory activity (IC, 31.27 ± 0.72 μg/mL). Three novel potent ACE inhibitory peptides were screened from the hydrolysate: LLRP (IC, 3.44 ± 0.19 μM), LHWPR (IC, 17.67 ± 1.53 μM), and RFLR (IC, 42.52 ± 7.50 μM). The peptides inhibited ACE activity via mixed-competitive (LLRP and RFLR) or competitive (LHWPR) mechanisms, forming stable complexes. Spectroscopic and molecular docking analyses revealed distinct binding modes for each peptide.
Hydroxytyrosol (HT), a potent natural antioxidant in olive oil, has broad food and nutraceutical applications, but sustainable, scalable production remains challenging. Here, we achieved efficient HT biosynthesis in us...Hydroxytyrosol (HT), a potent natural antioxidant in olive oil, has broad food and nutraceutical applications, but sustainable, scalable production remains challenging. Here, we achieved efficient HT biosynthesis in using glucose-glycerol dual carbon sources, combining metabolic engineering and rational enzyme design. We eliminated feedback inhibition, deleted competing pathways, and rationally mutated the rate-limiting enzyme HpaBC (S462A/M293Y), which enhanced catalytic efficiency by improving substrate entry, oxygen availability, and shortening reaction distance, as revealed by molecular dynamics simulations. Global cofactor optimization boosted HT titer to 4.90 g/L in shake flasks. Under optimized dissolved oxygen in a 5 L bioreactor, HT production reached 13.25 g/L, the highest reported titer in to date, providing a robust strategy for sustainable HT biomanufacturing.
Flowering time is a critical trait in Chinese cabbage, particularly in stalk-type cultivars. In this study, we characterized two allelic early flowering mutants and of Chinese cabbage whose mutant characteristics were...Flowering time is a critical trait in Chinese cabbage, particularly in stalk-type cultivars. In this study, we characterized two allelic early flowering mutants and of Chinese cabbage whose mutant characteristics were similar and insensitive to photoperiod, vernalization, and also GA treatments. Genetic analysis revealed that the early flowering trait was controlled by a single recessive nuclear gene. The causal gene, , was identified via MutMap and KASP techniques, and its function was further validated by sequencing analysis of allelic mutants and transient transformation. BrEPCR1 was a homologue of EPCR1, a polycomb-related protein involved in histone deacetylation and heterochromatin silencing. ChIP-qPCR revealed that H4K5ac levels decreased at /3 in the mutant. The lost part of BrEPCR1, the EPL domain, was necessary for the interaction between BrEPCR1 and BrMBD9. had the application potential to promote the early maturity in stalk type of Chinese cabbage.
Vardanega R, Lüdtke FL, Loureiro L
… +6 more, Fernández-Carrera A, Santos J, Venâncio A, Pinheiro AC, González-Fernández Á, Vicente AA
J Agric Food Chem
· 2026 Jul · PMID 42311044
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This study investigated whether coencapsulation of cannabidiol (CBD) and piperine (PIP) in food-grade nanostructured lipid carriers (NLCs) enhances the CBD oral bioavailability. NLCs containing long-chain fatty acids wer...This study investigated whether coencapsulation of cannabidiol (CBD) and piperine (PIP) in food-grade nanostructured lipid carriers (NLCs) enhances the CBD oral bioavailability. NLCs containing long-chain fatty acids were formulated with 1% CBD and 1% PIP using either a CBD isolate (CBD) or a CBD extract (CBD). Both systems showed high encapsulation efficiency and good stability over 28 days, with similar physicochemical properties. However, the CBD form strongly influenced digestion behavior: NLC-PIP-CBD exhibited high bioaccessibility for CBD (87 ± 5%), while NLC-PIP-CBD showed markedly lower values (13 ± 4%). Based on these results, NLC-PIP-CBD was evaluated in a mouse pharmacokinetic study against NLC-CBD and CBD in hemp seed oil. While NLC-CBD did not improve absorption, NLC-PIP-CBD doubled the CBD systemic exposure, confirming that PIP codelivery significantly enhanced oral CBD absorption.
Plasmalogens are ether phospholipids. They have a vinyl ether bond at -1 and polyunsaturated fatty acids at -2, and are abundant in marine- and animal-derived foods. Their unique structures may help maintain membrane org...Plasmalogens are ether phospholipids. They have a vinyl ether bond at -1 and polyunsaturated fatty acids at -2, and are abundant in marine- and animal-derived foods. Their unique structures may help maintain membrane organization, redox balance, and signaling. Human studies show that plasmalogen levels decline with age, and the drop is even sharper in Alzheimer's disease (AD), suggesting a link between dietary lipids and neurodegeneration. This narrative review summarizes dietary sources, molecular composition, and metabolism of plasmalogens, focusing on how structural diversity affects stability, bioavailability, and function. Based on cell and animal studies, we discuss potential mechanisms linking plasmalogens to AD pathology, including amyloid-β metabolism, oxidative stress, neuroinflammation, synaptic dysfunction, and myelin integrity. Unlike previous reviews, this work integrates structure-function relationships across food composition, lipid metabolism, and disease mechanisms. Finally, we discuss emerging uses in functional foods and precision nutrition, offering a fresh framework for dietary plasmalogens as part of neuroprotective strategies.
Sugar cane ( spp.) productivity is constrained by suboptimal growth and smut disease, underscoring the need for microbial strategies that integrate both plant growth promotion and disease suppression. Here, 59 sugar cane...Sugar cane ( spp.) productivity is constrained by suboptimal growth and smut disease, underscoring the need for microbial strategies that integrate both plant growth promotion and disease suppression. Here, 59 sugar cane-associated bacterial isolates were screened for both plant growth-promoting and antifungal activities; selected strains were further evaluated by biosafety assays, whole-genome sequencing, and untargeted LC-MS/MS metabolomics. The strain GXSB was prioritized as the lead candidate, and GXSA was included as a biologically relevant reference strain. Both strains inhibited , but GXSB exhibited stronger inhibition than GXSA (86.1% vs 81.3%) and was enriched in cyclic-nucleotide-associated metabolites. In greenhouse assays, GXSB reduced smut symptoms by 65.5%, increased plant height by 85%, and enhanced biomass by more than 3-fold relative to controls, with no detectable hemolytic activity. These findings identify GXSB as a promising dual-function candidate for further validation in promoting sugar cane growth and managing smut management.
The turnip mosaic virus (TuMV) is an RNA virus that poses a significant threat to crops. The coat protein (CP) plays a crucial role in the viral life cycle, including mediating host infection and cell-to-cell movement, w...The turnip mosaic virus (TuMV) is an RNA virus that poses a significant threat to crops. The coat protein (CP) plays a crucial role in the viral life cycle, including mediating host infection and cell-to-cell movement, which makes it an ideal target for antiviral drug development. In this study, a series of structurally diverse synthetic indolylureas were screened against TuMV CP using high-throughput methods such as isothermal titration calorimetry (ITC) and microscale thermophoresis (MST). This led to the identification of compound , which exhibits a high affinity for TuMV CP. Further experiments revealed that also binds to other homologous coat proteins from the genus, suggesting its potential as a broad-spectrum antiviral agent. Through molecular docking and point-mutation validation, the H183A residue in TuMV CP was identified as a key site for binding. In vivo experiments demonstrated that exhibits superior antiviral activity compared with ningnanmycin. Transmission electron microscopy observations indicated that disrupts the viral particle integrity. Additionally, omics analysis and defense enzyme activity assays showed that this compound was also found to modulate the plant immune responses. Together, these findings provide new insights into and experimental evidence for the development of novel plant antiviral strategies.