Search Results (1,453)

S-adenosyl-L-homocysteine hydrolase (SAHH) is an evolutionarily conserved enzyme present in eukaryotes, bacteria, and archaea. As the rate-limiting enzyme in the methionine cycle, it catalyzes the reversible hydrolysis of S-adenosyl-L-homocysteine (SAH) to adenosine and homocysteine, thereby modulating the S-adenosylmethionine/SAH ratio and cellular methylation potential. Dysregulation of SAHH activity is causally linked to cancer, cardiovascular disorders, and neurodegenerative conditions. This review systematically examines the biological distribution, catalytic mechanisms, structural architecture, and regulation of SAHH across diverse species. We highlight lineage-specific adaptations—including C-terminal truncation, a 40-residue substrate-binding-domain insertion, and a His-Phe molecular gate—that fine-tune substrate preference, cofactor affinity, and thermostability, with metal ions and NAD⁺ serving as key modulators of activity and conformational dynamics. These variations exemplify an evolutionary trade-off between catalytic efficiency and structural rigidity, particularly pronounced in archaeal and thermophilic orthologs. Collectively, these insights underpin the enzyme’s multifaceted translational value: SAHH serves as a therapeutic target for diverse diseases (e.g., cancer, viral infections, tuberculosis), a source of diagnostic/prognostic biomarkers (e.g., plasma homocysteine and SAH/SAM ratio), and a versatile biocatalyst for synthesizing pharmaceutical-grade adenosine and its derivatives. By integrating mechanistic, structural, and evolutionary perspectives, this review establishes a unified framework that explains these functional adaptations and their translational implications. This framework guides the rational development of SAHH-targeted inhibitors, diagnostic tools, and engineered biocatalysts, with broad applications in precision medicine and biotechnology. Full article

The ubiquitin–proteasome system (UPS) has traditionally been described as a tightly regulated degradative network driven mainly by the specificity of its ubiquitin-conjugating enzymatic components. The 26S proteasome is the catalytic arm of the system that acts downstream to the conjugation machinery. For a long time, it has been considered to be a constitutive multi-subunit proteolytic complex that recognizes in a non-discriminatory manner ubiquitin-marked target substrates with less than a handful of exceptions. However, emerging evidence reveals that the 26S proteasome function is also dynamically regulated by multiple factors, such as subunit composition and synthesis, post-translational modifications, and spatial localization, all of which are tightly regulated by the metabolic and stress states of the cell. Importantly, dysregulation of these newly emerging regulatory mechanisms has pathogenic sequelae. These mechanisms fine-tune proteasome activity and expand its role as an active regulator of protein homeostasis rather than being a passive degradation machinery. Given the rapid expansion of these findings and their impact on our understanding of proteasome biology, an integrated overview of these regulatory mechanisms is timely. Full article

Foodborne pathogens rely on colonization, biofilm formation, virulence expression, and environmental adaptation as fundamental biological drivers of food safety risk. Quorum sensing (QS), a cell-density-dependent microbial communication mechanism, coordinates the expression of these key phenotypes by integrating intraspecies, interspecies, and host-derived signals, making QS an attractive intervention target in food microbial control. Although QS research has advanced considerably in recent years, existing reviews have largely focused on individual bacterial species or specific classes of signal molecules. A systematic integration of how QS coordinately drives both food spoilage and pathogen virulence remains lacking. In this review, we conceptualize the QS network as a central regulatory hub connecting microbial signal perception to hazardous phenotype expression. We systematically examine the mechanistic roles of QS in food spoilage, biofilm formation, host colonization and invasion, and toxin production. We also summarize current QS-targeted intervention strategies, including inhibition of signal synthesis, enzymatic signal degradation, receptor antagonism, and indirect regulation via beneficial microorganisms. Building on the available evidence, we further analyze the key challenges limiting practical application: signal system specificity, ecological safety, industrial-scale feasibility, and microbial adaptability. Overall, QS-based strategies offer a non-bactericidal route for food microbial control, although substantial barriers remain for translation into complex food matrices. Reframing QS function and intervention from the perspective of food safety risk formation provides an analytical framework that bridges mechanistic understanding with practical application. This framework also establishes a theoretical foundation for developing next-generation food preservation and foodborne disease control strategies. Full article

The escalation of antimicrobial resistance (AMR) represents a serious global threat to public health, with AMR-associated mortality estimated to increase by 70% by 2050. As pathogens evolve through enzymatic inactivation, target modification, efflux-mediated clearance, biofilm formation, and broader genetic adaptation, conventional therapies are increasingly compromised, while the antibiotic development pipeline remains critically constrained by high discovery and development costs, weak commercial incentives, and the escalating complexity of resistance mechanisms. This review comprehensively synthesizes advanced pharmacological and biotechnological innovations designed to circumvent these entrenched resistance mechanisms. We highlight the development of novel therapeutic classes, particularly lariocidin, which disrupts bacterial protein synthesis via a previously unexploited ribosomal-binding site. Moreover, we critically evaluate molecular interventions, emphasizing CRISPR/Cas-based gene silencing and genome editing as precise tools to neutralize specific resistance determinants, such as the mecA gene in methicillin-resistant Staphylococcus aureus (MRSA). Concurrently, we explore the integration of engineered nanoparticles to revitalize existing antimicrobials by overcoming biofilm barriers, improving drug solubility, and enabling targeted delivery. Collectively, mastering the evolving AMR landscape requires a multidimensional framework that seamlessly integrates these novel molecular targets with advanced rapid diagnostics and robust international governance. Full article

Foods rich in polyphenols are known to promote oral health by modifying the enamel pellicle. In doing so, they reduce bacterial adhesion, biofilm maturation, and erosion. The goal of this study was to screen local herbal drugs available in Central Europe for their potential suitability as part of a diet promoting oral health by targeting the initial stages of biofilm formation. To achieve this, an in situ study was conducted to evaluate the effects of the four polyphenol-rich herbal extracts of blackcurrant leaves, oak bark, horse chestnut leaves, and sweet chestnut leaves on early bacterial adhesion and biofilm formation on tooth enamel over an 8 h period. This research aimed to identify natural remedies that could support oral hygiene by targeting the initial stages of biofilm formation. Study Design and Experimental Procedures: Aqueous extracts were prepared by ultrasonic extraction. Eight human subjects wore bovine enamel slabs intraorally for 8 h. After 1 min of pellicle formation, the subjects rinsed with 8 mL of the extracts for 10 min, followed by intraoral exposure without food. An 8 h-exposure without rinse served as the negative control; 0.2% chlorhexidine gluconate (CHX) served as the positive control. After 8 h, bacterial adhesion and biofilm matrix formation on the enamel slabs were quantified ex vivo using DAPI/Concanavalin A staining and fluorescence microscopy. The LIVE/DEAD™ BacLight™ assay was used to assess bacterial viability. Statistical analysis was performed by the Mann–Whitney U test and Kruskal–Wallis test (p < 0.05), as well as the Bonferroni–Holm correction (p < 0.01). Results and Conclusions: The screened herbal drugs did not demonstrate a statistically significant impact on the number of adherent bacteria, suggesting that their mode of action may not directly interfere with bacterial adhesion mechanisms. However, all four extracts exhibited consistent trends toward reduced glucan formation and decreased bacterial viability. The observed inhibition of glucan formation indicates that these drugs may potentially target the enzymatic pathways responsible for polysaccharide synthesis. By disrupting glucan production, the structural integrity of the biofilm matrix might be compromised, which indirectly affects bacterial survival within the biofilm environment. Full article

Hydrogen peroxide (H₂O₂) is a key reactive oxygen species involved in both cellular signaling and oxidative stress, making its reliable detection essential in biological and environmental systems. Electrochemical sensing has emerged as a promising approach for H₂O₂ monitoring due to its high sensitivity, rapid response, and suitability for in situ analysis. This review provides a comprehensive overview of nanostructured metal oxide electrodes for non-enzymatic electrochemical detection of H₂O₂. The effects of material composition, nanostructure morphology, and synthesis strategies (particularly hydrothermal methods) on sensor performance are critically discussed. Special attention is given to our previously reported studies, enabling a consistent comparison of structure–property relationships under similar experimental conditions. Furthermore, the application of these sensors in plant stress analysis is examined, including both the monitoring of oxidative stress and the evaluation of stress mitigation strategies using metal oxide nanoparticles. The role of nanoparticles as reactive oxygen species scavengers and enhancers of plant antioxidant systems is highlighted, demonstrating their ability to reduce H₂O₂ levels and improve plant physiological status under adverse environmental conditions. Overall, this work emphasizes the dual functionality of nanostructured materials as both sensing platforms and active agents for stress mitigation, highlighting their potential in agricultural and environmental applications. Full article

Biocatalysis has emerged as a mainstay in the field of sustainable chemical synthesis owing to its high selectivity, mild reaction conditions, and reduced environmental impact. Traditional enzyme engineering approaches, such as rational design and directed evolution, are often associated with limited throughput and a limited understanding of sequence–structure–function relationships, despite high experimental costs. In recent years, the integration of machine learning (ML) into enzyme engineering has emerged as a transformative approach, enabling data-driven prediction, design, and optimization of biocatalysts, thereby enhancing performance and applications. This review provides a comprehensive overview of ML-guided strategies to improve key enzymatic parameters, including the turnover number (k_cat), substrate affinity (K_m), and catalytic efficiency (k_cat/K_m), with a focus on mechanistic insights and performance outcomes. The integration of ML models into design–build–test–learn (DBTL) cycles accelerated directed evolution, reduced screening efforts, and enabled targeted mutagenesis. Beyond applications, this review also discusses the current limitations of ML-guided approaches, including data scarcity, model interpretability, and challenges in predicting complex mutations and allosteric effects. The gap between computational predictions and experimental outcomes is identified, and the role of ML integration with enzyme kinetics, molecular dynamics, and high-throughput experimentation is emphasized. Future directions, such as generative AI, explainable ML, and autonomous laboratories, are discussed for next-generation biocatalytic applications. Full article

Anti-nutritional factors (ANFs) in terrestrial plant feeds constrain efficient herbivore production, an issue intensified by rising feed costs and growing demand for animal products. Unlike previous reviews that focus on single ANFs or feed types, this review provides an integrated, cross-species framework linking ANF chemistry, rumen microbial interactions, and mitigation strategies. It examines major ANF classes—tannins, phytates, saponins, oxalates, protease inhibitors, lectins, glucosinolates, and gossypol—and their distribution and biochemical modes of action. Mechanistic pathways are grouped into digestive effects (reduced palatability and enzyme inhibition), microbial effects (altered rumen microbiota and fermentation), metabolic effects (impaired absorption), and mineral interactions (nutrient complexation and chelation). Species-specific responses are evaluated, emphasizing the partial detoxification capacity of the rumen microbiome and the dose-dependent nature of ANF effects. Mitigation strategies—physical, chemical, microbial, enzymatic, probiotic, and genetic—are critically assessed for efficacy, scalability, and sustainability. Emerging metabolomic and metagenomic evidence shows that certain ANFs confer functional benefits at controlled doses; for example, tannins improve nitrogen retention, saponins reduce methane, and phytic acid scavenges free radicals. This synthesis supports strategic management rather than complete elimination, informing safe and sustainable use of terrestrial feeds under evolving food-security and environmental challenges. Full article

Osteoarthritis (OA) is a chronic, debilitating degenerative joint disease whose prevalence is rising markedly with the rapid aging of the global population. In this study, we investigated the chondroprotective efficacy of NP-2007, an enzymatically hydrolyzed low-molecular-weight collagen from Cervi cornu, using IL-1β-stimulated SW1353 human chondrocyte cells and a medial meniscal transection (MMT)-induced OA rat model. In SW1353 cells, NP-2007 considerably suppressed the expression of inflammatory mediators (iNOS, COX-2) and cytokines (TNF-α, IL-6) without cytotoxicity. Crucially, it restored matrix homeostasis by downregulating catabolic enzymes (MMP-3, MMP-13, and ADAMTS-5) and upregulating anabolic markers (COL2A1, aggrecan), a process associated with the modulation of the Wnt/β-catenin and phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathways and the recovery of the master chondrogenic factor SOX9. These in vitro findings were consistent with the in vivo results from the MMT model, where oral administration of NP-2007 (50 and 200 mg/kg) for 8 weeks effectively preserved articular cartilage structure and proteoglycan content while markedly reducing serum levels of catabolic biomarkers, including MMP-13 and COMP. Collectively, our findings demonstrate that NP-2007 exerts potent chondroprotective effects by modulating the balance between cartilage degradation and synthesis, suggesting its potential as a therapeutic candidate for the management of OA. Full article

The goal of this study is to assess how dietary Atractylodes lancea ultrafine powder (AUP) affects crucian carp (Carassius auratus) growth, body composition, and hypoxia tolerance. A total of 630 crucian carp (13.21 ± 0.17 g) were fed diets containing 0.0%, 3.0%, 6.0%, 9.0%, 12.0%, 15.0%, and 18.0% AUP for 15 days. In crucian carp, dietary AUP was observed to improve protein content and decrease weight gain and fat content (p < 0.05). The ideal AUP addition level for fish was 13.04%, based on broken-line regression analysis of the protein concentration in fish bodies. The tissues and organs of crucian carp showed increased enzymatic antioxidant activities and non-enzymatic antioxidant levels, indicating that dietary AUP reduced the production of reactive oxygen species (ROS) and inhibited lipid oxidation (p < 0.05). This study shows that dietary AUP has a significant anti-obesity impact in fish, which is strongly linked to boosting the anabolism of protein and antioxidant defense as well as improving glucose metabolism to reduce glucose as the raw material for body fat synthesis. Additionally, under hypoxic conditions, the addition of AUP increased duration time (DT) and decreased crucian carp oxygen consumption rate (p < 0.05). The DT broken-line regression study verified that 6.05% is the ideal AUP level for fish. According to our research, dietary AUP strengthens fish’s resistance to hypoxia by modifying the metabolism of proteins and carbohydrates for energy production and enhancing antioxidant defense in respiration-related tissues and organs. In conclusion, AUP may be a natural way to reduce fish obesity and hypoxic stress. Full article

Background/Objectives: The corpus luteum (CL) in the dog is the sole source of progesterone (P4) during diestrus and pregnancy, making it a key regulator of reproductive function. However, robust and functionally validated in vitro models of canine luteal cells remain limited. This study aimed to establish and characterize a reproducible primary culture system of canine luteal cells with preserved steroidogenic activity and regulatory responsiveness. Methods: Ovaries containing CLs were collected from five clinically healthy bitches undergoing routine ovariohysterectomy (OHE). Luteal tissue was mechanically fragmented and enzymatically digested using collagenase type II. Primary cultures were established using an explant-based approach and maintained in Dulbecco’s Modified Eagle Medium/Ham’s F-12 nutrient mixture (DMEM/F12) or Roswell Park Memorial Institute medium 1640 (RPMI 1640) supplemented with 20% fetal bovine serum (FBS). Cellular morphology, proliferation, expression of steroidogenic markers—steroidogenic acute regulatory protein (STAR) and 3β-hydroxysteroid dehydrogenase type 1 (HSD3B1), P4 secretion, and responsiveness to forskolin stimulation were evaluated. Results: Cultured luteal cells exhibited stable attachment, proliferation, and a predominantly spindle-shaped morphology. Both media supported maintenance of a steroidogenic phenotype, while RPMI 1640 enabled enhanced proliferation, allowing expansion up to passage three and efficient cryobanking. Cells remained functionally active, secreting progesterone for up to 28 days in vitro. Forskolin stimulation increased progesterone secretion up to 2.7-fold, confirming preserved cyclic AMP-dependent steroidogenic responsiveness. Conclusions: The canine CL is a reliable source of functionally competent luteal cells, and the established culture system represents a physiologically relevant in vitro model. To our knowledge, this is the first functionally validated in vitro model of the canine CL. This platform enables controlled investigations of luteal function, endocrine regulation, and mechanisms of P4 synthesis, supporting its application in mechanistic and translational reproductive research. Full article

As the world moves toward a circular bioeconomy, starch nanoparticles (SNPs) have emerged as key components for sustainable development. Traditional production methods have historically relied on harsh acid treatments; however, their substantial environmental footprint has catalyzed a much-needed shift toward “green” chemistry. This review explores the rise of eco-friendly synthesis strategies—including high-power ultrasound, mechanical milling, nanoprecipitation, and enzymatic hydrolysis—and explains how these “clean” methods allow us to precisely define the nanoparticles’ properties. Furthermore, the functional applications of SNPs are analyzed, focusing on their role as reinforcing agents in biodegradable packaging, natural stabilizers in food emulsions, and encapsulation matrices for targeted nutrient delivery. By connecting recent breakthroughs, this work identifies technological synergy, the integration of physical and biological methods, as the most promising route to overcome current yield and scalability limitations. Finally, a future perspective is proposed, focusing on what is needed to move these innovations from the lab to industrial applications, ensuring they are safe, effective, and truly sustainable for the global food sector. Full article

Background: Bacterial adaptation to environmental and chemical stress involves coordinated, system-level responses collectively described as perturbome. Understanding conserved elements within core perturbomes may reveal strategic vulnerabilities for antimicrobial development. Methods: In this study, we implemented an integrative framework combining functional and comparative genomics, drug–target interactions and molecular docking to prioritize conserved stress-response targets in Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. Results: A total of 147 genes from previously defined core perturbomes were analyzed through interactome reconstruction and functional enrichment. Interactome and functional analyses revealed significant connectivity and functional clustering, primarily associated with molecule biosynthesis, translation, transcriptional regulation, and energy metabolism. Orthology-based comparative genomics identified six conserved orthogroups shared across at least two species, representing key stress-adaptive nodes including fatty acid synthesis initiation, metabolic stress buffering, transcription termination (Rho), ATP synthesis, peptidoglycan remodeling, and UDP-glucose-mediated envelope biosynthesis. Drug–target interaction analyses suggested that these conserved proteins are modulated by enzymatic inhibitors, metabolite analogs, or active-site competitors. Structural and docking analyses focused on a selected protein, FabF (β-ketoacyl-ACP synthase II) and confirmed catalytically coherent binding of cerulenin within the active site, with high concordance between experimentally resolved and AlphaFold-predicted structures, supporting the reliability of structure-based prioritization. Conclusions: Overall, the results demonstrate that bacterial stress responses converge on evolutionarily conserved metabolic and regulatory elements essential for homeostasis and tolerance to perturbations, being the first work integrating core perturbome data from different microorganisms. The proposed perturbome-informed framework provides a rational strategy to identify robust, broad-spectrum antimicrobial targets and highlights opportunities for drug repurposing and future experimental validation. Full article

This study focuses on the synthesis of glucovanillin mediated by UGT109A1 and its mechanism against Type 2 Diabetes Mellitus (T2DM). Recombinant UGT109A1 successfully synthesized glucovanillin from vanillin using UDP-Glc as the sugar donor. Through network pharmacology, 140 potential targets were identified. Seven key targets were further screened using LASSO and SVM-RFE algorithms. Among these, SLC5A1 and ADK showed strong diagnostic potential, with AUC values ranging from 0.85 to 0.89. Immune infiltration analysis linked these core targets to M2 macrophages. Single-cell transcriptomics revealed that ADK is widely expressed but enriched in B cells, while TLR9 is confined to plasmacytoid dendritic cells (pDCs). Cell-to-cell communication analysis identified a pDC-to-B cell signaling axis. In vitro assays demonstrated that glucovanillin exhibits concentration-dependent inhibitory activity against α-glucosidase with moderate potency, with an IC₅₀ of 413.84 ± 12.80 μM. Molecular docking, 200 ns molecular dynamics simulations (MD), and MM/PBSA calculations showed that glucovanillin binds more strongly to α-glucosidase (−7.4 kcal/mol) than vanillin (−5.4 kcal/mol). Therefore, the glycosylation mediated by UGT109A1 enhanced the bioactivity and targeting specificity of vanillin. In summary, glucovanillin exerts anti-T2DM effects through a dual mechanism involving α-glucosidase inhibition and regulation of key targets, making it a promising lead compound for T2DM treatment. Full article

Ionic liquids remain attractive alternatives as multifunctional media for the sustainable biosynthesis of lactones. Their unique physicochemical properties, including negligible vapor pressure, high thermal stability, and tunable polarity, offer significant advantages in terms of biocatalyst stabilization and reaction selectivity. For lactone synthesis, ionic liquids facilitate improved control over enzymatic transformations, enable efficient catalyst recycling, and reduce solvent consumption. This review summarizes recent advances in the application of ionic liquids as solvents or support modifiers in enzymatic lactone synthesis, focusing also on ε-caprolactone biosynthesis. A green chemistry metrics evaluation was also performed for selected examples from the literature. The role of ionic liquids in enhancing process efficiency and supporting green, sustainable process design is critically discussed, highlighting their potential for the development of more sustainable and environmentally friendly lactone production technologies. Full article