Search Results (193)

Proper nutrition is a fundamental determinant of human health and is structured through the intake of various nutritional components: (i) macronutrients, including carbohydrates, lipids, and proteins, which provide energy and essential structural materials for metabolic and physiological processes; (ii) micronutrients, such as vitamins and minerals, although required in smaller quantities, play crucial roles as enzymatic cofactors and regulators of numerous biochemical pathways; (iii) natural bioactive compounds (NBCs), substances found in plant-based foods (including polyphenols, carotenoids, phytosterols, and sulfur compounds) that exert protective effects thanks to their antioxidant and anti-inflammatory properties, contributing to the prevention of numerous chronic non-communicable diseases (CNCDs) [...] Full article

Organoselenium chemistry has undergone remarkable development over the past five decades, evolving from its initial association with high toxicity into a field with pivotal contributions to materials science, organic synthesis, catalysis, and Medicinal Chemistry. Among the diverse biological activities displayed by organoselenium compounds, their redox behaviour is particularly compelling, as many of these molecules act as efficient mimetics of the antioxidant enzyme glutathione peroxidase (GPx). In this work, we investigated the GPx-like activity of a series of N,N′-diaryl selenoureas toward the depletion of H₂O₂ and cumene hydroperoxide (CumOOH) as model ROS. Their reactivity was correlated with the electronic nature of the aryl substituents using a Hammett-type analysis, revealing a strong dependence of the reaction rate on remote electronic perturbations within the aromatic ring. Combined UV and NMR studies provided mechanistic evidence supporting a catalytic cycle in which selenoureas, operating at sub-stoichiometric loadings (1 mol%) and using a thiol as a cofactor-like molecule, can be used to efficiently scavenge ROS with half-lives of only a few minutes (~10–60 min). Furthermore, these selenoureas exhibited potent antiproliferative activity across several human solid tumour cell lines. Overall, these results offer mechanistic insight into the ROS-eliminating pathways of selenoureas and highlight their potential as chemopreventive or anticancer agents. Full article

Imine reductases (IREDs) have emerged as valuable biocatalysts for the asymmetric synthesis of chiral amines, key intermediates in numerous active pharmaceutical ingredients. Their ability to operate under mild reaction conditions with high chemo- and stereoselectivity provides an attractive alternative to conventional metal-catalyzed or chemical reduction processes. However, the broader industrial application of wild-type IREDs is often constrained by their limited substrate scope and moderate catalytic efficiency. Recent advances in biocatalysis have demonstrated that engineered IREDs can catalyze the reduction of a wide range of natural and non-natural imines, significantly expanding their applicability in pharmaceutical and fine chemical synthesis. In parallel, enzyme immobilization strategies have proven highly effective for improving operational stability, facilitating enzyme reuse, and enabling continuous flow biocatalytic processes. Efficient cofactor regeneration systems have further enhanced the practical implementation of IRED-based transformations. Advances in protein engineering, including structure-guided design, semi-rational mutagenesis, and directed evolution, have generated enzyme variants with improved catalytic activity, stereoselectivity, and substrate tolerance. The integration of high-throughput screening technologies and machine-learning-assisted enzyme design has further accelerated the discovery and optimization of efficient IRED biocatalysts. This review summarizes recent progress in the protein engineering and immobilization of IREDs and discusses future perspectives for their industrial application. Full article

Salivary aldehyde dehydrogenases (ALDHs), particularly ALDH3A1 and ALDH1A1, serve as frontline enzymatic defenses in the oral cavity, detoxifying reactive aldehydes generated through metabolic activity, microbial fermentation, and environmental exposures. These enzymes are essential for maintaining redox homeostasis, mucosal integrity, and immune modulation. However, under chronic metabolic stress, such as in diabetes, oral inflammation, and cancer, salivary ALDHs become vulnerable to non-enzymatic glycation by reactive carbonyl species like methylglyoxal. This modification impairs cofactor binding, catalytic activity, and structural stability, thereby compromising detoxification capacity at a time of heightened aldehyde burden. This review provides the first insights into ALDH glycation and particularly that of salivary ALDH, examining its structural mechanisms, disease-specific consequences, and emerging protective strategies. Special focus is given to natural compounds, including curcumin, thymoquinone, resveratrol, carnosine, and EGCG, that prevent glycation or restore ALDH function via carbonyl scavenging, antioxidant activation, and NAD⁺/SIRT1 pathway modulation. We also highlight critical research gaps, such as the absence of site-specific glycation maps, lack of salivary gland-based models, and limited availability of ALDH3A1-specific activators. Importantly, we propose that the glycation status of salivary ALDHs may serve as a non-invasive biomarker of oxidative stress and therapeutic response in metabolic and inflammatory disorders. By bridging biochemical insights with translational potential, this review establishes ALDH glycation as a mechanistic and clinically actionable axis in oral and systemic health. Full article

Natural pasture, the primary feed source in sheep production, often provides insufficient levels of essential minerals and vitamins required for proper metabolic regulation during pregnancy and early development. This study aimed to analyze patent developments of mineral and vitamin complexes (MVCs) for pregnant ewes and lambs and to evaluate the biochemical and molecular relevance of their components based on scientific evidence. A search of the World Intellectual Property Organization (WIPO) database using the keywords “vitamins for sheep” and “minerals for sheep” identified 120 patents related to sheep feed additives, including 23 specifically formulated for pregnant ewes and lambs. Comparative analysis revealed that calcium, selenium, iron, copper, cobalt, sodium, manganese, zinc, and vitamins A, D, and E were the most frequently included components. These micronutrients play critical roles in enzymatic activity, regulation of gene expression, antioxidant defense systems, and mineral homeostasis. In particular, zinc and selenium function as structural and catalytic cofactors for antioxidant enzymes such as superoxide dismutase and glutathione peroxidase, while vitamins A and D regulate cellular differentiation and calcium–phosphorus metabolism through transcriptional control mechanisms. Additionally, functional additives, including amino acids and plant-derived bioactive compounds, contribute to improved mineral bioavailability and modulation of metabolic pathways. The analyzed formulations demonstrate a consistent focus on correcting mineral deficiencies, enhancing antioxidant protection, and supporting metabolic adaptation during pregnancy and early postnatal development. Overall, the findings indicate that modern MVCs are rationally formulated to improve mineral utilization, physiological stability, and reproductive outcomes, highlighting their critical role in optimizing maternal health and offspring viability in sheep production systems. Full article

Bacterial poly(A) polymerases (PAPs) play an important role in RNA metabolism but remain poorly characterized outside Gammaproteobacteria. Here, we cloned and biochemically characterized the first PAP from Alphaproteobacteria, specifically from Marinobacter lipolyticus (Mli PAP). Using homology-based screening against E. coli PAP-1, we identified Mli PAP, sharing 54.8% sequence identity with its E. coli counterpart. The enzyme was expressed in E. coli but formed insoluble inclusion bodies; the active enzyme was purified as a fusion protein with the DsbA protein and used for functional assays. Mli PAP exhibited optimal activity at 30 °C and similar thermostability to E. coli PAP-1. ATP was the preferred substrate, with K_m comparable to E. coli PAP-1 (1.61 mM and 1.70 mM, respectively), and Mg²⁺ (10 mM) was identified as the optimal cofactor. Mli PAP displayed salt-dependent activity, with the most effective polyadenylation in KCl and inhibition by NaCl and ammonium salts, contrasting with the halophilic nature of its host. This study provides the first functional insights into PAPs from Alphaproteobacteria, broadening the understanding of PAP diversity and biochemical properties, as well as the potential applications of PAPs in biotechnology. Full article

Laccases are environmentally friendly biocatalysts capable of oxidizing a broad range of organic pollutants by using molecular oxygen as the sole electron acceptor, producing water as the only by-product. The cofactor-independent activity makes them attractive for sustainable wastewater treatment, particularly for the removal of synthetic dyes. In this study, laccase from Trametes versicolor was evaluated for the decolorization of the azo dye Methyl Orange, with emphasis on the effects of redox mediators and metal ions. Laccase alone exhibited maximum activity at pH 3.0, achieving 40.5% decolorization after 24 h. The addition of redox mediators markedly enhanced dye removal. Both synthetic mediators (ABTS, HBT, and TEMPO) and natural mediators (p-coumaric acid, vanillin, gallic acid, ascorbic acid, and syringaldehyde) improved decolorization in a concentration-dependent manner. Among them, ABTS and syringaldehyde were the most effective, achieving 98.8% and 96.9% decolorization, respectively, at 0.2 mM after 24 h, with syringaldehyde offering the advantage of natural origin. Metal ions also modulated laccase activity, with several ions enhancing decolorization at 0.5 mM, whereas higher concentrations were generally inhibitory; potassium ions showed the strongest enhancement (80.3%). Molecular docking analysis suggested favorable binding of Methyl Orange within the laccase active pocket, supported by predicted hydrogen-bonding and hydrophobic interactions. The molecular docking analysis was performed using a representative T. versicolor laccase structure and provides supportive computational insight into plausible enzyme–dye interactions rather than isoform-specific mechanistic validation. Overall, these findings demonstrate that optimization of mediators and metal ion concentrations significantly improve laccase-mediated dye decolorization, while docking provides supportive computational insight into possible enzyme–dye interactions relevant to sustainable wastewater treatment. Full article

Innovative developments of GC-MS over the last two decades made this methodology a powerful tool for profiling a broad range of volatile metabolites and non-volatile ones of non-polar, semi-polar and even polar nature after appropriate derivatization. Indeed, the high potential of GC-MS in the analysis of low molecular weight metabolites involved in essential cellular functions (energy production, metabolic adjustment, signaling) made it the method of choice for the life and plant scientists. However, despite these advances, due to their intrinsic thermal lability, multiple classes of hydrophilic low-molecule weight metabolites (like nucleotides, sugar phosphates, cofactors, CoA esters) are unsuitable under the high-temperature conditions of the split–splitless (SSL) injection and GC separation, which makes the analysis of such compounds by GC-MS challenging. Therefore, to ensure comprehensive coverage of the plant metabolome, the GC-MS-based metabolomics platform needs to be efficiently combined with other metabolomics techniques and instrumental strategies. Moreover, to get a deeper insight into dynamics of plant cell metabolism in response to endogenic and exogenic clues, integration of the metabolomics data with the output obtained from other post-genomics techniques is desired. Therefore, here, we overview different strategies for the integration of the GC-MS-based metabolite profiling output with the data, acquired by other metabolomics techniques in terms of the multi-platform metabolomics approach. Further, we comprehensively discuss the implementation of the GC-MS-based metabolomics in multi-omics strategies and the data integration strategies behind this. This approach is the promising strategy, as it gives deep and multi-level insight into physiological processes in plants in the systems biology context, with consideration of all levels of gene expression. However, multiple challenges may arise in the way of integrating data from different omics technologies, which are comprehensively discussed in this review. Full article

Due to the neuroprotective and antioxidant properties, phenylethanoid glycosides (PhGs) are valuable plant-derived compounds. Traditional extraction methods are constrained by low yields and limited resources, prompting the integration of synthetic biology and enzyme engineering technologies for sustainable production. This review summarizes the advances in the microbial synthesis of PhGs, emphasizing the elucidation of biosynthetic pathways, enzyme engineering modifications of glycosyltransferases and acyltransferases, and strategies for optimizing microbial cell factories in Escherichia coli and Saccharomyces cerevisiae. Significant advancements encompass the efficient synthesis of verbascoside and echinacoside in S. cerevisiae, as well as the comprehensive elucidation of the echinacoside biosynthetic pathway in Cistanche spp., including the identification of key steps catalyzed by a rhamnosyltransferase, a CYP450 hydroxylase, and a terminal glucosyltransferase that enable pathway reconstruction in S. cerevisiae. We conduct a systematic analysis of methods to address the biosynthetic bottlenecks via protein engineering, including rational design and directed evolution, as well as the metabolic engineering strategies such as precursor enhancement and cofactor recycling. Additionally, we investigate the synthesis of non-natural PhG analogues and the prospective integration with AI-assisted design, emphasizing the significant potential of microbial systems in overcoming the supply challenges for medicine-food homologous ingredients. Full article

UVB radiation induces cutaneous damage through oxidative stress and immune dysregulation. This study investigated the therapeutic potential of Gastrodia elata fermented by Lactobacillus salivarius AACE1 (GL) in a mouse model of UVB-induced skin injury. Results demonstrated that GL treatment significantly improved skin morphology, enhanced antioxidant activities (SOD and GSH), reduced oxidative damage (MDA), and balanced inflammatory mediators by upregulating TGF-β and IL-10 while downregulating TNF-α, IL-6, and IL-1β. Transcriptomic analysis revealed that GL specifically activated NOD-like receptor signaling pathway components (Nlrp3, Casp4, and Gbp2/5) while inducing Tnfaip3 to establish negative feedback control. Metabolomic profiling confirmed that fermentation transformed the metabolite landscape, enriching collagen-related dipeptides, antimicrobial/anti-inflammatory metabolites, and antioxidant cofactors. Importantly, comparative analysis showed that GL is more effective than vitamin E in coordinating multiple signaling pathways and maintaining inflammatory homeostasis. These findings establish GL as an effective natural product that alleviates UVB-induced skin damage through synchronized metabolic remodeling and controlled immune activation. Full article

Vitamin B1 (thiamine) is a water-soluble B vitamin. As a cofactor of many enzymes, it is essential for the proper functioning of many body systems and organs, including metabolic and energy metabolism. In extreme cases, vitamin B1 deficiency causes neurodegenerative disorders, including beri-beri, or cognitive impairment resulting from encephalopathy. B1 avitaminosis may result from increased demand, dietary errors, malabsorption, or excessive loss. Thiamine supplementation is used in cases of vitamin B1 deficiency or for preventative measures in situations of increased demand. Vitamin B1 can be administered enterally or parenterally (intravenously, intramuscularly, subcutaneously). The route and dose depend on the individual patient’s clinical situation. Hypersensitivity to vitamin B1 is rare and appears to be primarily associated with rapid intravenous infusion of large doses of thiamine hydrochloride over a short period (intravenous bolus). Hypersensitivity to thiamine administered by routes other than intravenous or intramuscular injection appears to be an incidental phenomenon. Thiamine should also be considered as an occupational allergen. The mechanism of thiamine hypersensitivity has not been clearly elucidated. However, considering the clinical nature and dynamics of the reaction, the most likely reaction seems to be an immediate type of hypersensitivity reaction (immunoglobulin E (IgE)-dependent), in which thiamine (but not its metabolites) acts as a hapten. Diagnosing hypersensitivity to vitamin B1 is difficult due to the lack of validated tests for additional testing. In individuals requiring thiamine supplementation who have experienced hypersensitivity to intramuscular or intravenous administration of this vitamin, switching to oral administration may be considered (provided this does not reduce treatment efficacy). This form of supplementation is usually well tolerated by individuals allergic to parenteral thiamine. However, if enteral supplementation does not guarantee the maintenance of therapeutic potential, thiamine desensitization may be considered, which seems to be an effective therapeutic method in such a clinical situation. Full article

Copper and nickel ions play pivotal, albeit distinct, roles as essential trace elements in living systems, and primarily serve as co-factors for a range of enzymes. However, as with all trace metal ions, excessive concentrations can exert adverse toxicological properties. Interestingly, the incorporation of these in cell culture media can establish novel chemical interactions, with their speciation status markedly influencing characteristics, including cell maturation, and cellular uptake mechanisms. Thus, the primary objective of this study was to investigate and determine the speciation status (i.e., complexation) of nickel(II) and copper(II) ions by biomolecules present in RPMI 1640 mammalian cell culture medium using virtually non-invasive high-resolution proton NMR analysis, an investigation of much relevance to now routine studies of their toxicological actions towards cultured cells. Samples of the above aqueous culture medium were ¹H NMR-titrated with increasing added concentrations of 71–670 µmol/L Ni(II)(aq.), and 0.71–6.7, 7.1–67 and 71–670 µmol/L Cu(II)(aq.), in duplicate or triplicate. ¹H NMR spectra were acquired on a JEOL ECZ-600 spectrometer at 298 K. Results demonstrated that addition of increasing concentrations of Ni(II) and Cu(II) ions to the culture medium led to the selective broadening of a series of biomolecule resonances, results demonstrating their complexation by these agents. The most important complexants for Ni(II) were histidine > glutamine > acetate ≈ methionine ≈ lysine ≈ threonine ≈ branched-chain amino acids (BCAAs) > asparagine ≈ aspartate > tyrosine ≈ tryptophan, whereas for Cu(II) they were found to be histidine > glutamine > phenylalanine ≈ tyrosine ≈ nearly all remaining aliphatic metabolites (particularly the wealth of amino acids detectable) > 4-hydroxyphenylacetate (trace culture medium contaminant), in these orders. However, Cu(II) had the ability to influence the linewidths of these signals at much lower added levels (≤7 µmol/L) than that of Ni(II), the broadening effects of the latter occurring at concentrations which were approximately 10-fold greater. Virtually all of these added metal ion-induced resonance modifications were, as expected, reversible on addition of equivalent or excess levels of the chelator EDTA. From this study, changes in the co-ordination sphere of metal ions in physiological environments can give rise to marked modifications in their physicochemical properties (e.g., redox potentials, electronic charges, the potential catalytic generation of reactive oxygen species (ROS), and cell membrane passages). Moreover, given that the above metabolites may also function as potent hydroxyl radical (^●OH) scavengers, these findings suggest that generation of this aggressively reactive oxidant directly from Cu(II) and Ni(II) ions in physiologically-relevant complexes may be scavenged in a ‘site-dependent’ manner. This study is of further relevance to trace metal ion research in general since it enhances our understanding of the nature of their interactions with culture medium biomolecules, and therefore provides valuable information regarding their overall chemical and biological activities, and toxicities. Full article

Overweight and obesity are major public health concerns among children and adolescents worldwide. The most prevalent form is exogenous–constitutional obesity, which is driven by a sedentary lifestyle and an unhealthy diet in which caloric intake exceeds energy expenditure. Beyond their association with chronic disease, these factors are closely linked to deficits in cognitive development and executive functions essential for learning (including working memory, sustained attention, planning, behavioral self-regulation, and cognitive flexibility). Oxidative stress (OS), characterized by the accumulation of reactive oxygen species (ROS) in cells and extracellular fluids, is a significant potential mediator in childhood obesity and an important contributor to its comorbidities. The antioxidant defense system (AOD)’s activity largely depends on levels of trace element cofactors, which determine the body’s resistance to adverse environmental factors (the “maladaptation phenomenon”). OS and trace element deficiencies contribute to the development of morphological changes in the brain, thus serving as a critical connecting link between childhood obesity and cognitive impairment. Non-pharmacological interventions are the most accessible and effective approach for prevention and treatment. Bioactive compounds derived from food and natural plants, classified as antioxidants and phytopreparations, may represent a promising complementary approach. These compounds are most effective when used in combination with sustained lifestyle modifications in children. Research in this area can help define future directions for study and develop targeted intervention strategies in the pediatric population. The aim of this review is to examine the relationship between OS, antioxidant cofactor micronutrients, and cognitive outcomes in childhood obesity and to explore mechanisms, evidence, and therapeutic opportunities. Full article

Background/Objectives: Inborn errors of amino acid metabolism (IEAAMs) are inherited disorders caused by defects in amino acid catabolism, biosynthesis, or transport. In this review, we aimed to synthesise recent evidence on the clinical manifestations and current and future therapeutic strategies for major IEAAMs. Methods: A narrative review was undertaken on studies published up to November 2025. No fixed start date was set. Instead, earlier studies were included if historically significant or frequently cited in contemporary guidelines, and emphasis was placed on recent developments over the last 5–10 years. Evidence was identified through structured searches of PubMed, clinical trial registries, and public communications on selected IEAAMs, which were synthesised in textual and tabular form. Results: Management across IEAAMs involves the restriction of amino acids or natural proteins, disease-specific dietary formulations, micronutrient optimisation, cofactor or enzyme replacement, and pharmacological chaperones. This is supported by structured monitoring and emergency regimens to prevent catabolic crises. Organ transplantation remains crucial for select indications, such as liver transplantation in hereditary tyrosinaemia with liver disease. Novel approaches include substrate reduction, the pharmacological targeting of upstream pathways, viral vector gene transfer, and liver-directed mRNA therapy. Several of these novel approaches have entered clinical trials, but many remain in the preclinical stage. Conclusions: Despite advances in the treatment of IEAAMs, many patients still experience significant morbidity. Future focus should be on further refining emerging molecular and gene-based treatments and optimising neuroprotective and metabolic targets. The equitable implementation of personalised, life-spanning treatments within multidisciplinary rare disease services will be essential. Full article

Human DNA dioxygenase ABH2 is a key enzyme of the AlkB family of Fe(II)/α-ketoglutarate-dependent oxygenases, which is specialized in removing alkyl groups from damaged DNA bases in the cell nucleus. At the same time, the occurrence of single-nucleotide polymorphisms (SNPs) in the human ABH2 gene can lead to amino acid substitutions that, in turn, may disrupt the normal functioning of the ABH2 enzyme. Currently, databases contain information about more than 2500 nucleotide substitutions in the ABH2 gene. Using a comprehensive bioinformatics approach, in this review, we analyzed over 200 non-synonymous ABH2 SNPs with eleven prediction programs to identify variants capable of negatively affecting its enzymatic activity. The combination of various programs with different evaluation algorithms and scoring approaches allows us to more reliably identify potentially deleterious amino acid substitutions. Moreover, the differences between the programs used allowed for comparison of their tendency to predict amino acid substitutions as deleterious. Structural analysis of the ABH2-substrate complex showed that selected functionally significant SNPs often affect the organization of the active site, reduce the efficiency of substrate binding, and/or disrupt the coordination of Fe²⁺ and α-ketoglutarate cofactors, leading to changes in catalytic efficiency. The data obtained from the conducted analysis suggest that naturally occurring polymorphisms in the ABH2 gene found in the human population may reduce the repair efficiency of DNA dioxygenase ABH2 and, consequently, modulate susceptibility to oncogenesis and influence the effectiveness of antitumor therapy for carriers of these SNPs. Full article