d-Tagatose is a rare hexose sugar with excellent properties, and its synthesis catalyzed by d-tagatose 4-epimerase (T4E) represents a competitive novel pathway. In this study, T4E derived from the was screened and syste...d-Tagatose is a rare hexose sugar with excellent properties, and its synthesis catalyzed by d-tagatose 4-epimerase (T4E) represents a competitive novel pathway. In this study, T4E derived from the was screened and systematically characterized. By reshaping the microenvironment of the active pocket, mutant M3(S131D/H410W/T279S) was constructed, which showed a 3.89-fold higher conversion rate compared with the wild-type (WT) enzyme. Kinetic parameter analysis and molecular dynamics (MD) simulations revealed that M3 had enhanced substrate affinity, hydrogen bond network, charge properties, and channel accessibility. Finally, the conversion rates of d-fructose to d-tagatose catalyzed by the purified M3 enzyme and M3 whole-cell catalysts reached 29.46% and 26.2%, respectively. Additionally, the dual-enzyme cascade reaction of M3 with glucose isomerase (GI) TEGI-M-L38M-V137L was constructed, achieving a 13.16% yield of d-tagatose from d-glucose. This study demonstrates that T4E-M3 is a promising biocatalyst for d-tagatose production, laying the foundation for its subsequent industrial application.
Alginate lyases are important for producing bioactive oligosaccharides and combating biofilms, demonstrating significant application potential in medicine, food, industry, and other sectors. However, few cold-adapted en...Alginate lyases are important for producing bioactive oligosaccharides and combating biofilms, demonstrating significant application potential in medicine, food, industry, and other sectors. However, few cold-adapted enzymes with high thermostability exist. Here, we identified a novel alginate lyase, PpAly7A (26.75% sequence identity), which adapts to various pH, temperatures, salt concentrations, metal ions, and surfactants. PpAly7A effectively disrupts PAO1 biofilms (36.51% removal) and exhibits exceptional thermostability (half-life 4.39 days at 50 °C). Site-directed mutagenesis and molecular dynamics simulations revealed that synergistic interactions among disulfide bonds, proline residues, salt bridges, and hydrogen bond networks maintain thermostability by enhancing global stability and compactness. Notably, the E180Q mutation further extends the half-life to 7.25 days while retaining most catalytic activity, demonstrating a favorable activity-stability trade-off. This study deepens the understanding of the structure-function relationships of enzymes and provides a viable molecular template for the rational design of thermostable industrial enzymes.
poses a major threat to global soybean production. Here, CRISPR/Cas9-mediated promoter replacement revealed that the V-ATPase A subunit gene () plays an important role in the growth, development, and pathogenicity of , h...poses a major threat to global soybean production. Here, CRISPR/Cas9-mediated promoter replacement revealed that the V-ATPase A subunit gene () plays an important role in the growth, development, and pathogenicity of , highlighting its potential as a molecular target. Using this target, a multistage docking-based virtual screening identified compound as a potent antioomycete inhibitor, with EC values of 4.082-8.974 μg/mL against six oomycetes. markedly inhibited mycelial growth and key developmental stages in and protected soybean leaves from infection without affecting soybean seed germination or seedling growth. Mechanistically, potentially interacted with PsVHA-A, inhibited V-ATPase activity, and disrupted vacuolar homeostasis, cell wall structure, and energy metabolism. Collectively, our findings establish VHA-A as a promising oomycete-specific fungicidal target and identify as a valuable lead compound for the development of selective antioomycete agents.
4-Hydroxyphenylpyruvate dioxygenase (HPPD) is a key target for new herbicide development. To explore novel and effective HPPD inhibitors for weed control, a series of pyrazole-benzothiazine hybrids was designed and synth...4-Hydroxyphenylpyruvate dioxygenase (HPPD) is a key target for new herbicide development. To explore novel and effective HPPD inhibitors for weed control, a series of pyrazole-benzothiazine hybrids was designed and synthesized by combination and enol tautomerism strategies. Bioassays indicated that compound strongly inhibited HPPD (HPPD), with an IC of 0.06 M, which was 8 and 3.7 times stronger than topramezone and mesotrione. Additionally, compound demonstrated over 80% herbicidal activity against the tested weeds at 150 g a.i./ha, with only 10% injury to maize, peanut, and cotton. Molecular docking revealed that compounds and formed bidentate chelation with Co and π-π interactions with Phe381 and Phe424 of HPPD (PDB: 7CQS), and stable hydrogen bonds with Asn282. Microscale thermophoresis (MST) and fluorescence titration (FT) confirmed their strong binding affinity to HPPD, with Asn282 as a potential binding site. Thus, pyrazole-benzothiazine hybrids are a promising scaffold for HPPD inhibitor development.
This study employed AlCl/ZnCl bimetallic salt catalysis with ethanol organic solvent for sugar cane bagasse pretreatment. The process retained 80.12% cellulose while removing ∼70% xylan and 80.56% lignin, achieving 90.18...This study employed AlCl/ZnCl bimetallic salt catalysis with ethanol organic solvent for sugar cane bagasse pretreatment. The process retained 80.12% cellulose while removing ∼70% xylan and 80.56% lignin, achieving 90.18% glucose conversion during enzymatic hydrolysis. Comprehensive characterization and DFT calculations revealed a synergistic dual-action mechanism: strong Lewis acid Al extensively cleaved β-O-4 bonds (reducing them to 40.68%), whereas weak Lewis acid Zn interacted broadly with side chains and aromatic rings. This synergy facilitated substantial lignin removal and minimized total phenolic OH content in residual lignin (1.32 mmol/g), thereby suppressing unproductive cellulase adsorption. Additionally, the bimetallic system exhibited stronger overall interactions with the model compound GG compared to single-component systems. This integrated approach establishes a robust foundation for efficient lignocellulose fractionation and downstream bioconversion.
Agricultural practices are increasingly shifting toward more sustainable and regenerative approaches; however, their impact on crop metabolism and associated biological effects remains incompletely understood. In this st...Agricultural practices are increasingly shifting toward more sustainable and regenerative approaches; however, their impact on crop metabolism and associated biological effects remains incompletely understood. In this study, H NMR-based metabolomics combined with functional biological assays was used to investigate metabolic modulation in blueberry, cauliflower, and peach extracts obtained under regenerative and conventional agricultural systems. Metabolic profiling revealed clear, matrix-dependent differences in carbohydrates, amino acids, organic acids, and phenolic-related compounds between cultivation strategies. Multivariate analysis (PLS-DA) showed discrimination between regenerative and conventional samples. Cauliflower exhibited the most pronounced metabolic modulation, followed by peach, whereas blueberry showed more selective changes. Functional assays showed enhanced fibroblast migration in peach extracts, increased cytoprotective and detoxifying responses in cauliflower, and antiangiogenic activity in blueberry samples. Overall, these results suggest that agricultural practices may influence the metabolic and functional properties of plant-derived foods and demonstrate the value of combined metabolomic and biological approaches for the comparative evaluation of regenerative production systems.
Fenoxaprop--ethyl (FE), an acetyl-CoA carboxylase (ACCase) inhibitor, disrupts fatty acid synthesis, but downstream lipid disturbances remain unknown. Here, we performed integrative physiological, enzymatic, lipidomic, a...Fenoxaprop--ethyl (FE), an acetyl-CoA carboxylase (ACCase) inhibitor, disrupts fatty acid synthesis, but downstream lipid disturbances remain unknown. Here, we performed integrative physiological, enzymatic, lipidomic, and transcriptomic analyses to investigate FE-induced lipid disruption in rice seedlings. Lipidomics revealed that FE decreased total lipid content by ∼35%, mostly reducing free fatty acids, phosphatidylglycerol, and lysophosphatidylethanolamine, particularly long-chain (≥C16) species. KEGG enrichment identified fatty acid elongation as a key pathway, with 32 DEGs predominantly downregulated. Most genes for elongation enzymes (KCS, KCR, HCD, ECR) were downregulated, and six key DEGs were confirmed by qRT-PCR. Enzyme assays showed time-dependent inhibition of these enzymes, peaking at 53-66% on day 7 post-treatment. Multiomics analysis suggested that ACCase inhibition disrupts fatty acid elongation, depleting critical lipids. We propose that FE disrupts lipid homeostasis via "cascade inhibition of lipid metabolism", where suppression of fatty acid elongation amplifies phytotoxicity.
To address the escalating pesticide resistance in lepidopteran pests, a 1,2,4-oxadiazole motif was integrated into an isoxazoline scaffold to synthesize 25 novel derivatives. Compound exhibited exceptional insecticidal...To address the escalating pesticide resistance in lepidopteran pests, a 1,2,4-oxadiazole motif was integrated into an isoxazoline scaffold to synthesize 25 novel derivatives. Compound exhibited exceptional insecticidal activity against (LC = 0.21 mg/L), significantly outperforming both fluxametamide (LC = 1.36 mg/L) and ethiprole (LC = 11.2 mg/L). Furthermore, demonstrated 36-fold greater efficacy against than ethiprole. Mechanistic investigations utilizing two-electrode voltage-clamp electrophysiology revealed that functions as a potent antagonist of the 1 GABA receptor, effectively suppressing GABA-evoked currents. Molecular docking and molecular dynamics simulation indicated that binds deeply within the receptor pore, establishing stable interactions with critical residues (e.g., Ala302 and Phe206) to disrupt channel gating. This study presents a successful structural optimization strategy for isoxazoline insecticides and highlights as a promising candidate for next-generation pest management.
Hexavalent chromium (Cr[VI]) is a significant environmental contaminant that poses substantial risks to agricultural safety and public health. Although Cr(VI)-induced hepatic injury is associated with oxidative stress an...Hexavalent chromium (Cr[VI]) is a significant environmental contaminant that poses substantial risks to agricultural safety and public health. Although Cr(VI)-induced hepatic injury is associated with oxidative stress and organelle dysfunction, the crosstalk between ionic homeostasis disruption and organelle-specific pathophysiology remains incompletely elucidated. Herein, we identified significant suppression of peroxiredoxin 1 (PRDX1) following Cr(VI) exposure, concomitant with intracellular calcium depletion, mitochondrial lipid peroxidation, endoplasmic reticulum (ER) stress, and ferroptosis. Mechanistically, Cr(VI)-driven PRDX1 deficiency initiates calcium-mediated mitochondrial oxidative damage and ER stress, synergistically triggering the ferroptosis cascade that culminates in hepatic injury. These findings establish PRDX1 as the master regulator of Cr(VI) hepatotoxicity and uncover a novel calcium-dependent axis that converges mitochondrial redox imbalance with ER proteotoxic stress to drive ferroptotic cell death. This study provides fundamental insights into the toxicological role of PRDX1 in Cr(VI) exposure and advances theoretical frameworks for understanding chromium-associated pathophysiological mechanisms.
The fall armyworm, , is a polyphagous pest that poses significant economic threats to a large scale of high-value crops and exhibits high resistance to synthetic insecticides, potentially impacting nontarget organisms. T...The fall armyworm, , is a polyphagous pest that poses significant economic threats to a large scale of high-value crops and exhibits high resistance to synthetic insecticides, potentially impacting nontarget organisms. To mitigate undesirable ecological impacts, novel triazole derivatives were synthesized in an eco-friendly manner to develop highly effective insecticides. Toxicological bioassays revealed insecticidal performance alongside larvae in their third larval instar. The investigated hybrids revealed LC values ranging from 1554 to 2588 mg/L. Notably, compound displays remarkable insecticidal potency (LC = 1554 mg/L). DFT calculations and molecular docking simulations provided insights into key features that override the bioactivity of the synthesized triazole hybrids. A molecular docking study was performed using PDB IDs 2ACE and 4COF, showing that binary triazole 9 has the highest binding affinity for the essential amino acids, consistent with experimental assessments. Also, in vivo biochemical and histological analyses revealed substantial disorders in the investigated hybrid.
PF08 is a significant food spoilage organism belonging to a species widely utilized for agricultural biocontrol, yet the post-translational regulation of its biofilm formation remains poorly understood. This work explore...PF08 is a significant food spoilage organism belonging to a species widely utilized for agricultural biocontrol, yet the post-translational regulation of its biofilm formation remains poorly understood. This work explored the role of lysine succinylation (Ksucc) in regulating CsrA activity, a global post-transcriptional regulator. NanoHPLC-MS/MS identified Lys38 (K38), a highly conserved residue within the CsrA RNA-binding interface, as a functional succinylation site. Site-directed mutagenesis revealed that K38 succinylation significantly inhibits biofilm formation by reducing c-di-GMP levels, exopolysaccharide production, and surface hydrophobicity, while enhancing flagellar-mediated motility. Colonization assays on foods, contact surfaces, and the rhizosphere confirmed that CsrA succinylation restricts biofilm formation across diverse ecological niches. These results identify K38 succinylation of CsrA as a regulatory modification that modulates the transition between motile and sessile lifestyles in PF08. These findings expand the understanding of post-translational control of biofilm formation in this ecologically versatile bacterium relevant to food spoilage and agricultural environments.
Analysis of pesticide residues in guttation fluids remains challenging due to the small quantity of fluids. A multimode inlet technique was utilized to increase the detectability of 12 pesticide residues in guttation flu...Analysis of pesticide residues in guttation fluids remains challenging due to the small quantity of fluids. A multimode inlet technique was utilized to increase the detectability of 12 pesticide residues in guttation fluids on leaves collected with a pipet. A 500-μL sample of guttation fluid was vortex-extracted with an equal volume of dichloromethane. The extracts were dried over sodium sulfate anhydrous, followed by gas chromatography-tandem mass spectrometry analysis. Multiple injection mode and solvent vent mode were combined for a large-volume injection (e.g., 10 μL). The method was validated for the analysis of 12 systemic pesticides in guttation fluids for detection selectivity (no interferences), method limit of quantification (1-5 μg/L), linearity (>0.993), accuracy (70.7%-116%), precision (2.0%-17.0%), and matrix effect (-3.9%-304%). Kinetics of pesticide residues in the guttation fluids over time were monitored. This method is hopefully useful for the analysis of pesticide residues in guttation fluids.
Arctigenin, a lignan ingredient from burdock ( L.), has therapeutic promise against ulcerative colitis. Here, we reveal its mechanism based on pyroptosis of colonic macrophages. The data showed that arctigenin specifical...Arctigenin, a lignan ingredient from burdock ( L.), has therapeutic promise against ulcerative colitis. Here, we reveal its mechanism based on pyroptosis of colonic macrophages. The data showed that arctigenin specifically inhibited N-terminal fragment of gasdermin D (GSDMD-N)-mediated pyroptosis of colonic macrophages in mice with dextran sulfate sodium salt-induced colitis. Mechanistically, arctigenin activated estrogen receptor β (ERβ), which bound to the intron region of carnitine palmitoyl transferase 1A (CPT1A) to down-regulate CPT1A expression, thereby reducing fatty acid oxidation (FAO) in macrophages. This reduction in FAO inhibited GSDMD-N acetylation and oligomerization, ultimately attenuating pyroptosis. In colitis mice, overexpression of CPT1 or knockout of ERβ abolished the protective effects of arctigenin. These findings demonstrate that arctigenin exerts anticolitis effect through activating ERβ and down-regulating FAO to prevent GSDMD-N oligomerization and consequent pyroptosis of colonic macrophages. Burdock and its main active ingredient arctigenin merit development as functional foods or therapeutic agents for ulcerative colitis.
False-positive annotations in untargeted lipidomics limit biological interpretation. A modified equivalent carbon number (mECN) rule was developed with independently fitted parameters to overcome the fixed coefficient li...False-positive annotations in untargeted lipidomics limit biological interpretation. A modified equivalent carbon number (mECN) rule was developed with independently fitted parameters to overcome the fixed coefficient limitation of the classical ECN model, enabling quantifiable false-positive filtering. Validated with a fish oil of known composition and five independent data sets, this workflow improved precision by 8.4% over traditional methods, with 0.5% recall loss and 3.4% F1 gain. Multilevel verification integrating structural (subclass, lipid species, fatty acid) and analytical (metabolite, pathway, network) dimensions was established to strengthen biological interpretation. Applied to a neuropathic pain model, six lipids (CE 20:5, LPC 20:5, LPE 20:5, PG 22:6_22:6, PE 16:1_22:6, and LPI 22:6) were associated with pain modulation, implicating fatty acid elongation/desaturation and phospholipase A2-mediated acyl chain remodeling. This study provides a robust analytical framework for untargeted lipidomics and mechanistic insight into ω-3 nutritional intervention for pain management.
Naturally occurring inhibitors of membrane-bound enzymes offer therapeutic potential for metabolic disorders such as obesity and fatty liver disease. Sphingomyelin synthase (SMS), which converts ceramide to sphingomyelin...Naturally occurring inhibitors of membrane-bound enzymes offer therapeutic potential for metabolic disorders such as obesity and fatty liver disease. Sphingomyelin synthase (SMS), which converts ceramide to sphingomyelin, is a promising target in this regard. In this study, -9-docosenoic acid (9Z-DSA) was identified from the dietary seaweed () as a natural SMS inhibitor. Inhibition assays using HeLa cell lysates expressing SMS1 and SMS2 a reliable model for membrane enzyme studies were analyzed by liquid chromatography-mass spectrometry (LC-MS). The nonpolar hexane extract of exhibited SMS inhibitory activity, and purification revealed 9Z-DSA (47% purity) as the active compound, with IC values of 4.5 ± 1.3 μM for SMS1 and 5.8 ± 1.8 μM for SMS2. Molecular docking and dynamics simulations indicated strong binding interactions between 9Z-DSA and the SMS active sites. These findings demonstrate that 9Z-DSA functions as a natural SMS inhibitor with potential as a nutraceutical candidate for metabolic disease management.
Identifying new leads is vital for agrochemical innovation against . Unprecedented combinations of classic benzimidazole fungicides with prevalidated antifungal heterocycle-oxazolines were designed for novel scaffolds. A...Identifying new leads is vital for agrochemical innovation against . Unprecedented combinations of classic benzimidazole fungicides with prevalidated antifungal heterocycle-oxazolines were designed for novel scaffolds. A modular [3 + 2] cyclization of -amino -heterocycles with 3-bromo-2-oxopropanoate enabled the efficient construction of 6 distinct heterocycles, facilitating access to novel imidazole-oxazolines for unbiased antifungal phenotypic screening. 2-(Imidazo[1,2-]pyridazin-2-yl)oxazoline was identified as a novel antifungal hit. Guided optimization delivered promising candidates (- (), -, ) active against thiabendazole-resistant (F.70). Notably, compound exhibited an EC of 0.4 μM (0.15 mg/L) against F.70, demonstrating >200-fold greater potency than thiabendazole and promising activity against carbendazim-resistant strains BM-1 and BM-2. It could disintegrate the mycelium of as detected by SEM. Photostability, improved synthesis, divergent modifications, cheminformatic simulation, and biosafety evaluation of this candidate were also investigated. The unique structural and biological profile of this oxazoline scaffold facilitates antifungal agrochemical discovery.
The high comorbidity of obesity and depression causes serious public health events when systemic inflammation induced by disturbed immunity and gut microbiota is supposed to be involved. Previously, the promoting effect...The high comorbidity of obesity and depression causes serious public health events when systemic inflammation induced by disturbed immunity and gut microbiota is supposed to be involved. Previously, the promoting effect of triticale bran insoluble dietary fiber (TIDF) on anti-inflammatory bacteria besides inhibiting proinflammatory bacteria was uncovered with bound phenolics (BPs) in it of great importance, while the potential effects of TIDF on obesity-concomitant depression have not been reported yet. This study found that TIDF treatment could relieve depression-like behavior, systemic inflammation, and Th17/Treg imbalance in obese mice. The regulation of microbiota-derived SCFAs was closely involved. Besides down-regulating harmful bacteria while boosting and by both TIDF and its BPs-removed counterpart (T-D), BPs enhanced the microbiota-modulating effects of TIDF on specific SCFAs-producing genera, including and . Our study was deemed to promote the application of food nutritional factors in relieving obesity-concomitant emotional disturbance.
Photopharmacology provides a strategy for green pesticide design. Starting from tolfenpyrad, azobenzene-modified photochromic ligands were synthesized. showed high bioactivity and photoisomerization efficiency but limi...Photopharmacology provides a strategy for green pesticide design. Starting from tolfenpyrad, azobenzene-modified photochromic ligands were synthesized. showed high bioactivity and photoisomerization efficiency but limited / activity difference. To enhance this, terminal phenyl modifications yielded , which displayed amplified photoregulatory activity differences across pests. Notably, exhibited a 4.8-fold photoinduced activity enhancement against larvae and a 6.6-fold photoinduced activity reduction against , while achieved a 35-fold reduction against . Molecular docking and binding free energy calculations indicated stronger -isomer binding to the target via multiple hydrogen bonds. Molecular orbital analysis revealed a minimal HOMO-LUMO gap difference (0.05 eV) between isomers, suggesting activity differences arise from conformational changes in charge distribution rather than energy variations. This work demonstrates a rational design approach for high-performance photoresponsive pesticides.
The transition from growth to survival mode is an important strategy for plants to cope with extreme environments. However, the molecular regulatory network governing this process has not yet been fully elucidated. In th...The transition from growth to survival mode is an important strategy for plants to cope with extreme environments. However, the molecular regulatory network governing this process has not yet been fully elucidated. In this study, we reveal that Brassinosteroid (BR) negatively regulates drought tolerance in a concentration-dependent manner, whereas Abscisic acid (ABA) enhances drought resistance by antagonizing BR. Drought tolerance assays confirm that the transcription factor cold and drought resistance 1 (OsCDR1) is essential for ABA-enhanced drought resistance via suppression of BR signaling. OsCDR1 directly targets and activates the transcription of , and through physical interaction with dwarf and low-tillering (DLT), alleviates the inhibitory effect of DLT on , thereby negatively regulating BR signaling. Our findings reveal a novel ABA-OsCDR1-BR regulatory module that governs drought responses in rice.