Orange-fleshed (O) potato tubers are rich in nutritionally beneficial carotenoids. However, the metabolic and regulatory mechanisms underlying their accumulation remain incompletely understood. Metabolomic profiling iden...Orange-fleshed (O) potato tubers are rich in nutritionally beneficial carotenoids. However, the metabolic and regulatory mechanisms underlying their accumulation remain incompletely understood. Metabolomic profiling identified 32 carotenoids and showed that the O genotype reached peak carotenoid accumulation at the early swelling stage, with 17 xanthophylls present at higher levels than in a white-fleshed (W) genotype. Transcriptome analysis revealed that five biosynthetic genes, including (), were significantly upregulated in O genotype, whereas () was more highly expressed in the W genotype. Bulk segregation analysis of a backcross population identified five quantitative trait loci, including and , as key genetic determinants of carotenoid variation. Integrated coexpression networking and molecular validation indicated that and likely regulate and expression, respectively. Our findings elucidate the metabolic, genetic, and transcriptional mechanisms controlling carotenoid deposition in tubers, providing a foundation for genetic improvement of potato quality.
Conventional d-tagatose production via isomerization suffers from thermodynamic equilibrium limitations. An alternative two-step pathway employing aldose reductase and galactitol 2-dehydrogenase enables thermodynamically...Conventional d-tagatose production via isomerization suffers from thermodynamic equilibrium limitations. An alternative two-step pathway employing aldose reductase and galactitol 2-dehydrogenase enables thermodynamically driven conversion of d-galactose to d-tagatose through a galactitol intermediate. In this study, a novel galactitol 2-dehydrogenase from sp. L1K22 (WP_208553890.1) was identified and characterized. The recombinant His-tagged enzyme, purified by nickel-affinity chromatography, displayed peak activity at pH 9.5 and 35 °C. It showed specific activities of 36.7 U/mg toward galactitol and 48.8 U/mg toward d-sorbitol. Kinetic analysis for galactitol revealed a of 13.4 mM, of 1,889.0 min, and catalytic efficiency (/) of 141.0 mM min. When expressed in a galactitol-producing chassis via pET-22b (+), this enzyme drove efficient d-tagatose biosynthesis, attaining a titer of 8.80 g/L. These findings establish the sp. L1K22 enzyme as an effective biocatalyst for thermodynamically favorable d-tagatose production.
Seed priming presents a promising strategy for regulating plant growth and development by using nanomaterials. However, the response of plant roots and their associated microorganisms to nanopriming remains poorly unders...Seed priming presents a promising strategy for regulating plant growth and development by using nanomaterials. However, the response of plant roots and their associated microorganisms to nanopriming remains poorly understood. In this study, carbon dots (CDs) were synthesized and applied for rice seed priming. The results showed that CD priming significantly enhanced photosynthesis and growth in rice seedlings. Notably, CDs improved root architecture, increased cell wall thickness and lignification, enhanced root vitality, stimulated Ca influx and K efflux, and upregulated genes related to aquaporins and stress resistance. Microbial analysis revealed that CD priming significantly enriched microbial communities involved in nitrogen transformation and absorption while reducing pathogenic species. Consequently, these effects induced by CD priming on rice seedlings conferred significant resistance to salt, 2,4-D, and 2,4-D-Na stresses. This study provides a sustainable approach to promoting plant growth and mitigating environmental stress.
Cocoa butter is the main raw material for chocolate and other food products. However, cocoa butter production is constrained by land availability and environmental factors. Oleaginous yeasts offer a promising alternative...Cocoa butter is the main raw material for chocolate and other food products. However, cocoa butter production is constrained by land availability and environmental factors. Oleaginous yeasts offer a promising alternative. In this study, 103 yeast strains were isolated and screened for their lipid accumulation capacity. Among them, MYL-1 produced high-level cocoa butter equivalent (CBE), accounting for 45.3% of total triacylglycerols. Under nitrogen limitation with an optimal C/N ratio of 80:1, MYL-1 achieved 58.2% fatty acids of dry cell weight, 14.9 g/L biomass, and 8.7 g/L fatty acid titer. Fed-batch cultivation in a 5-L bioreactor yielded 24.6 g/L lipids with a productivity of 0.17 g/L/h, and a balanced C16:0/C18:0 ratio near 1:1, enhancing the suitability for CBE production.
Plant biomass, predominantly composed of lignocellulose, exhibits structural and chemical complexity that limits its biological deconstruction. This review evaluates the oxidative strategies utilized by microorganisms, p...Plant biomass, predominantly composed of lignocellulose, exhibits structural and chemical complexity that limits its biological deconstruction. This review evaluates the oxidative strategies utilized by microorganisms, primarily filamentous fungi, to overcome the recalcitrance. We integrate recent experimental breakthroughs with structural bioinformatics to map the expanding landscape of redox enzymes, including newly discovered AA families. Central enzymatic systems involved in lignin depolymerization include laccases and lignin-active peroxidases, whereas polysaccharide-degrading oxidoreductases comprise lytic polysaccharide monooxygenases (LPMOs) and their auxiliary redox partners. Hydrogen peroxide acts as a central intermediate that bridges enzymatic sources with sinks, subject to spatiotemporal regulation. The review also incorporates the synergistic interplay between oxidoreductases and nonenzymatic radical systems, offering refined mechanistic insights. These understandings underpin the design of next-generation biorefineries, accelerating the transition toward a sustainable circular bioeconomy.
(-)-Epigallocatechin has multiple bioactivities. ANR catalyzes its biosynthesis, with unclear structure and mechanism restricting enzyme modification. Here, we first screened ANRs from 21 plant species and identified AN...(-)-Epigallocatechin has multiple bioactivities. ANR catalyzes its biosynthesis, with unclear structure and mechanism restricting enzyme modification. Here, we first screened ANRs from 21 plant species and identified ANR (TgANR) as the most efficient candidate based on molecular docking and in vitro assays. Through integrated alanine scanning and computational prediction, we constructed a combinatorial mutant N15S/F106E/P182S, which exhibited a 3.88-fold higher EGC synthesis activity than the wild-type enzyme. The interaction mechanism between TgANR and the substrate delphinidin as well as the structure-activity relationship in the catalytic production of EGC were studied on the basis of protein engineering modification, thus clarifying the catalytic mechanism of TgANR. Furthermore, transient overexpression of the mutant gene in kernels significantly increased EGC accumulation. This study not only elucidates the structure activity relationship of TgANR but also provides an efficient enzyme variant and a protein engineering framework for boosting EGC biosynthesis in plants.
fucoidan's complex, heterogeneously sulfated structure hinders the preparation of well-defined fuco-oligosaccharides (FOs). While enzymatic depolymerization offers a mild and selective route, efficient enzymes specifical...fucoidan's complex, heterogeneously sulfated structure hinders the preparation of well-defined fuco-oligosaccharides (FOs). While enzymatic depolymerization offers a mild and selective route, efficient enzymes specifically tailored for fucoidan remain scarce. Here, we identified and characterized OUC-MgFucD1, a novel GH107 family fucoidanase from, exhibiting unprecedented substrate specificity. OUC-MgFucD1 exclusively cleaves α-(1→3)-linked regions of fucoidan, producing sulfated mono-, di-, and tetrasaccharides. Its hydrolysis pattern is strictly modulated by local sulfation motifs. To enhance industrial applicability, a consensus-based rational design approach integrating three complementary platforms was employed to develop a stabilized variant, OUC-MgFucD1-H359D. This engineered enzyme exhibits exceptional thermal robustness, retaining over 60% activity after 12 h at 40 °C, whereas the wild-type enzyme undergoes complete inactivation. Our findings underscore the functional diversity of GH107 family, providing a robust molecular framework and an efficiency enzymatic tool for the precision-tailored valorization of fucoidan into FOs.
In recent years, the harm caused by mycotoxins to people cannot be ignored, and agricultural products contaminated by them are often highly toxic to humans. In this context, the requirements for the detection of mycotoxi...In recent years, the harm caused by mycotoxins to people cannot be ignored, and agricultural products contaminated by them are often highly toxic to humans. In this context, the requirements for the detection of mycotoxins in food production have gradually improved. Metal-organic frameworks (MOFs) offer the advantages of structural design flexibility and integrated functional synergies, enabling them to satisfy the stringent material specifications for the aforementioned nanomaterial sensing technologies. This paper summarizes the latest research on MOF-driven sensors (MDsensors) for early mycotoxin detection in food, covering MOF merits, synthesis techniques, sensing mechanisms, enhancement strategies, and potential applications in food safety early warning systems. Notably, this paper emphasizes the performance merits of MOFs and the working principles of MDsensors, subsequently summarizing their applications in detecting mycotoxins to enhance food safety. Finally, it looks ahead to the prospects of integration with technologies such as portable devices and artificial intelligence.
Obesity is a serious global health concern driven by unhealthy diets. Lipophilic grape seed proanthocyanidin (LGSP) has demonstrated potential lipid-lowering properties, yet its underlying mechanisms remain incompletely...Obesity is a serious global health concern driven by unhealthy diets. Lipophilic grape seed proanthocyanidin (LGSP) has demonstrated potential lipid-lowering properties, yet its underlying mechanisms remain incompletely elucidated. This study investigated the antihyperlipidemic effects of LGSP in high-fat diet (HFD)-fed mice. LGSP supplementation attenuated HFD-induced obesity, organ hypertrophy, fat accumulation, liver injury, and hepatic inflammation. Gut microbiota analysis revealed that LGSP significantly increased microbial abundance and diversity in HFD-fed mice and positively reshaped the gut flora structure by decreasing the Firmicutes/Bacteroidetes ratio. Integrated serum metabolomic and liver transcriptomic analyses identified key regulators involved in lipid metabolism, including palmitoylcarnitine, hydrocortisone, acitretin, embelin, and 1-(11Z-eicosenoyl)-glycero-3-phosphate, as well as the genes Serpina3n and Acnat2. Further validation confirmed that LGSP enhances bile acid metabolism, promotes thermogenesis, and improves energy metabolism. These findings underscore the potential of LGSP as an effective functional food component for managing hyperlipidemia.
Cocoa flavanols show promise for inhibiting type 2 diabetes (T2D). The components responsible for this remain poorly understood. We compared cocoa extract (CE) supplementation against matched epicatechin (EC) or a high-m...Cocoa flavanols show promise for inhibiting type 2 diabetes (T2D). The components responsible for this remain poorly understood. We compared cocoa extract (CE) supplementation against matched epicatechin (EC) or a high-molecular-weight fraction (HMW) in a T2D mouse model induced by high-fat (HF) plus streptozotocin (STZ). Mice were fed HF diet with STZ alone or with CE, EC, or HMW. HF + STZ animals fed CE had blunted weight gain and reduced insulin resistance compared to HF + STZ control as measured by the homeostasis model assessment of insulin resistance. However, these improvements are driven by reduced fasting glucose and insulin, as peripheral insulin resistance was not observed in the model when accounting for the fasting glucose. HF + STZ-induced dysbiosis was partially reversed by CE and HMWs. These findings demonstrate the beneficial effects of CE on the gut microbiome and cardiometabolic markers, which were not fully achieved by EC or HMW, suggesting synergistic effects of flavanols.
Seafood provides important nutrients to our diets, particularly omega-3 polyunsaturated fatty acids (PUFAs). Compared with other common seafood, sea snails represent an underutilized resource that may be exploited to hel...Seafood provides important nutrients to our diets, particularly omega-3 polyunsaturated fatty acids (PUFAs). Compared with other common seafood, sea snails represent an underutilized resource that may be exploited to help meet global PUFA demands. However, sea snails exhibit complex fatty acid (FA) profiles, making an accurate and detailed characterization a considerable analytical challenge. By employing an advanced mass spectrometry (MS) technique, i.e., covalent adduct chemical ionization (CACI), we characterized the FA profiles of five commonly consumed sea snail species. A straightforward method was established to distinguish monounsaturated fatty acid methyl esters (FAMEs) from isobaric dimethyl acetals (DMA), verifiable by retention-time models. FAME double-bond positional isomers were further identified by CACI-MS/MS diagnostic ions. Sea snails are abundant in plasmalogens at 82-333 mg/100 g food and long-chain omega-3 PUFA up to 174 mg/100 g food. Therefore, sea snails, particularly saltwater varieties, show promise as mainstream seafood because of their high nutritional values.
A rapid method for producing -()-lutein esters from all--lutein esters was developed using I, AlCl, and sulfides as catalysts. -Isomer-rich lutein (38.6%) and -isomer-rich lutein esters (34.8%) by I were obtained. The an...A rapid method for producing -()-lutein esters from all--lutein esters was developed using I, AlCl, and sulfides as catalysts. -Isomer-rich lutein (38.6%) and -isomer-rich lutein esters (34.8%) by I were obtained. The antioxidant activities of -isomer-rich lutein and -isomer-rich lutein esters were higher than those of their all--isomers by Ferric Reducing Antioxidant Power, ORAC-L, and ABTS assays. The UV-A and UV-B blocking activities of -isomer-rich lutein esters were higher than those of all--lutein esters. After short-term ingestion by rats, the in vivo absorption concentration of -isomer-rich lutein (222.3 pmol/mL) in a high dose was higher than those of other forms of lutein (<125.6 pmol/mL). Enrichment of -lutein (-ratio: 59.5%) and -lutein esters (-ratio: 44.4%) was achieved in hexane and ethanol, respectively. We conclude, for the first time, that -isomer-rich lutein esters play a more important role than all--lutein esters in preventing oxidation and UV-blocking activity and are estimated to exhibit higher bioavailability than all--lutein esters and all--lutein.
Resveratrol (RSV) is a plant-derived polyphenol that has attracted attention for its potential antiobesity effects. However, the molecular mechanisms underlying its metabolic regulation remain unclear. In this study, an...Resveratrol (RSV) is a plant-derived polyphenol that has attracted attention for its potential antiobesity effects. However, the molecular mechanisms underlying its metabolic regulation remain unclear. In this study, an integrated network pharmacology analysis was combined with in vivo experiments to investigate the potential mechanisms associated with RSV-mediated metabolic improvement in high-fat diet (HFD)-induced obesity. The network pharmacology analysis suggested that RSV targets pathways related to lipid metabolism and energy homeostasis. In HFD-fed mice, RSV supplementation reduced body weight gain, improved glucose tolerance and lipid profiles, and suppressed adipose inflammation. RSV treatment was also associated with increased β3-AR, phosphorylated AMPKα, and UCP1 expression and was accompanied by corresponding changes in markers associated with lipolysis, fatty acid oxidation, and adipose thermogenesis. These results suggest that the antiobesity effects of RSV may be associated with β3-AR/AMPKα-related thermogenic signaling modulation, supporting its potential application as a food-derived bioactive compound for functional food development.
The tuber is the main edible and economic organ of potato ( L.). However, the molecular mechanisms underlying tuberization, particularly the crosstalk of phytohormones and transcription factors, remain inadequately under...The tuber is the main edible and economic organ of potato ( L.). However, the molecular mechanisms underlying tuberization, particularly the crosstalk of phytohormones and transcription factors, remain inadequately understood. In this study, we identified two AP2/ERF transcription factors, and , which are highly expressed in stolons and upregulated with swelling. Overexpression of significantly increased the number of stolons and tubers, ultimately enhancing yield. Integrated RNA-seq and DAP-seq analyses revealed that StRAP2.7a/b modulates the expression of multiple -zeatin (tZ) biosynthesis genes and tuberization-regulatory genes. It was observed that StRAP2.7a/b promotes tZ accumulation in stolons while repressing expression, thereby inducing stolon and tuber formation. Additionally, molecular dissection demonstrated that StRAP2.7a/b directly binds to and activates the promoter of , which is finely modulated by its interacting protein StCOL1. These findings advance our understanding of the mechanisms of tuberization and provide valuable genetic resources for breeding high-yielding potato cultivars.
The small brown planthopper, , exhibits escalating buprofezin resistance, yet the underlying molecular mechanisms remain unclear. In this study, we established a highly resistant near-isogenic line and utilized bulk segr...The small brown planthopper, , exhibits escalating buprofezin resistance, yet the underlying molecular mechanisms remain unclear. In this study, we established a highly resistant near-isogenic line and utilized bulk segregant mapping to localize a major resistance locus on chromosome 2. This led to the identification of a G932C target site mutation in chitin synthase 1 (), which was found at high frequencies in field populations. Molecular docking indicated that this mutation induces steric hindrance, disrupting buprofezin binding within the transmembrane chitin translocation channel. Additionally, synergism assays, transcriptome analyses, and RNAi confirmed that the overexpressions of cytochrome P450 and ATP-binding cassette transporter collectively confer metabolic resistance. Notably, significant cross-resistance between buprofezin and pymetrozine was observed. These findings demonstrate a dual mechanism of target-site and metabolic resistance to buprofezin in , providing crucial molecular markers for resistance monitoring and guidance for sustainable insecticide application strategies.
Robust and reproducible proteome coverage remains a major challenge in plant bottom-up proteomics because of tissue-specific biochemical complexity and variability during sample preparation and mass spectrometry (MS) acq...Robust and reproducible proteome coverage remains a major challenge in plant bottom-up proteomics because of tissue-specific biochemical complexity and variability during sample preparation and mass spectrometry (MS) acquisition. Here, we systematically evaluated four sample preparation workflows (in-solution digestion, FASP, S-trap, and SP3) combined with three MS acquisition strategies, including data-dependent acquisition (DDA), data-independent acquisition (DIA), and ion mobility-enabled four-dimensional DIA (4D-DIA), across rice leaves, roots, and seeds. All workflows generated comparable extraction profiles across tissues; however, SP3 exhibited lower median coefficients of variation (10.5 to 15.1%) while requiring substantially reduced processing time. Notably, 4D-DIA identified 3-fold to 4-fold more proteins than DDA in each tissue and modestly outperformed conventional DIA, with improved detection of low-abundance proteins and high quantitative reproducibility. Collectively, SP3 with the 4D-DIA workflow provides a sensitive, reproducible, and time-efficient platform for plant bottom-up proteome profiling, particularly for samples with limited material or high metabolite interference.
Siamenoside I (SI), a high-intensity sweetener, is limited in natural abundance. A β-glycosidase (CoExg1) from L7 (isolated from the rhizosphere) exhibits intrinsic sequential regioselectivity, enabling efficient SI pr...Siamenoside I (SI), a high-intensity sweetener, is limited in natural abundance. A β-glycosidase (CoExg1) from L7 (isolated from the rhizosphere) exhibits intrinsic sequential regioselectivity, enabling efficient SI production from mogroside V (MGV). Fermentation optimization increased SI yield from 70.34% (7 d) to 84.06% (72 h). CoExg1 rapidly cleaves the C-3 β-1,6 bond (89.35% SI in 20 min), then slowly hydrolyzes the C-24 bond to mogroside IIIE (MIIIE, 12 h)─a time-controlled process requiring no external regulation. Kinetic analysis showed a ∼50-fold higher catalytic efficiency toward MGV than toward SI. Molecular docking and dynamics revealed that the C-3 glycan of MGV binds in a catalytically competent conformation with stable hydrogen-bond networks, whereas SI's C-24 glycan hardly forms a productive binding mode, explaining the sequential selectivity. This work provides a sustainable SI production platform and unveils the structural basis of sequential regioselectivity in glycoside hydrolases.
Serine proteases from species are widely used in the feed industry, but their poor tolerance to high acidity and temperature limits their digestive efficacy in animals. To overcome this, we engineered the serine proteas...Serine proteases from species are widely used in the feed industry, but their poor tolerance to high acidity and temperature limits their digestive efficacy in animals. To overcome this, we engineered the serine protease SprD1 from D1 via structure-guided site-directed mutagenesis, using protein folding free energy (ΔΔ) analysis and loop flexibility modification. Multiple single and five combinatorial mutants were designed, among which the double mutant Mut5 showed the highest improvements. Mut5 retained 60 kU/mL activity at pH 2.0, about 3.3-fold higher than wild-type SprD1 (18 kU/mL), and maintained 36.4 kU/mL after 12 h at pH 2.2, demonstrating outstanding acid resistance. It preserved an ∼10% increase in activity after 4 h at 50-60 °C. In simulated gastric digestion of animal feed, Mut5 significantly enhanced amino acid release, with enzyme dosage and substrate concentration identified as key efficiency determinants. Overall, Mut5 is a high-performance protease combining acid stability, heat resistance, and superior feed hydrolysis.
The whitefly, , a globally significant pest transmitting over 200 plant viruses, has developed substantial resistance to insecticides, particularly neonicotinoids. Using near-isogenic lines, bulked segregant analysis (BS...The whitefly, , a globally significant pest transmitting over 200 plant viruses, has developed substantial resistance to insecticides, particularly neonicotinoids. Using near-isogenic lines, bulked segregant analysis (BSA), and genetic linkage analysis, we identified a major resistance locus on chromosome 6. The locus harbored dual mutations in the nicotinic acetylcholine receptor (nAChR) β1 subunit gene associated with high-level neonicotinoid resistance. Bioassays using gene-edited strains revealed that the R79E mutation conferred significant resistance to neonicotinoids. A field population carrying the R79E single mutation and exhibiting high-level resistance was subsequently identified using an optimized hybridization and backcrossing strategy combined with BSA. Furthermore, a six-year resistance monitoring program revealed that R79E is a major contributor to high-level neonicotinoid resistance in field populations. These findings improve our understanding of target-site resistance mechanism in agricultural pests and support the development of molecular tools for resistance monitoring and management.