Enzyme Activity Research Articles

The Role of Plant-Mycorrhizal-Fungal Interactions in Soil Health and Carbon Sequestration in Agroecosystems

Abstract: Understanding the intricate interactions between plants, mycorrhizal fungi, and soil health is crucial for sustainable agriculture and climate change mitigation. This review synthesizes current research to elucidate the multifaceted role of plantmycorrhizal-fungal interactions in soil health and carbon sequestration in agroecosystems. A meta-analysis of 75 studies spanning diverse ecosystems and plant species reveals a significant positive correlation between mycorrhizal colonization and soil organic carbon content, with an average increase of 25% compared to non-mycorrhizal systems. Furthermore, mycorrhizal symbiosis enhances soil structure and aggregation, promoting water retention and nutrient cycling, which are essential for maintaining soil fertility and resilience to environmental stressors. The contributions of different mycorrhizal types (arbuscular, ectomycorrhizal, and ericoid) to soil carbon dynamics are also examined, highlighting their unique roles in carbon allocation and stabilization. Additionally, the review discusses the potential implications of plant-mycorrhizal-fungal interactions for mitigating greenhouse gas emissions through enhanced carbon sequestration in agricultural soils. Insights into the mechanisms underlying these interactions, including mycorrhizal-mediated changes in root exudates, microbial communities, and soil enzymatic activities, are discussed. Overall, this review underscores the importance of integrating mycorrhizal symbiosis into agroecosystem management practices to enhance soil health, carbon sequestration, and climate resilience.

Species-Dependent Response of Brassica chinensis L. to Elevated CO2 Gradients Influences Uptake and Utilization of Soil Nitrogen, Phosphorus and Potassium

Employing elevated CO2 (eCO2), similar to increasing atmospheric CO2 in a greenhouse, is a common practice used to increase vegetable crop yields. However, the responses of nutrient availability, nutrient uptake and use efficiency in leafy vegetables to eCO2 remain largely unknown. The plant biomass production, nitrogen (N), phosphorus (P) and potassium (K) contents, nutrient uptake, and soil enzymatic activities of three Brassica chinensis varieties of ‘Longpangqing’, ‘Heimeiren’ and ‘Qingjiangbai’ were thus assessed under ambient CO2 (C0, 420/470 ppm, day/night) and eCO2 (C1, C0 + 33.3%C0; C2, C0 + 66.7%C0; and C3, C0 + 100%C0) for 4 months. Biomass production was significantly enhanced in these three tested B. chinensis varieties along with eCO2 gradients, with the highest biomass production under C3. Moreover, the eCO2 significantly increased plant N and K uptake, P and K use efficiency, soil saccharase, urease, and phosphatase activities, but decreased plant P content, soil available N and K, and catalase activity. Under eCO2, ‘Qingjiangbai’ exhibited higher biomass production and P and K uptake, without a decrease in soil available N and P. Consequently, more external fertilizer supplementation (especially N, followed by K) is required to maintain or promote Brassica chinensis yield under eCO2, while these management strategies must be carefully considered for different crop varieties.

Chemical Profile, Antioxidants, and Enzymatic Activity of the Kombucha Tea Beverages as a Potential Anti-obesity Beverage

This study aimed to investigate the chemical profile, antioxidant activity, and enzyme inhibition of black tea kombucha and kombucha without tea, as well as the relationship between these three parameters. Both kombuchas of different fermentation times (1, 7, 14, 21 days) were analysed. The pH and total soluble solids (TSS) of both samples decreased gradually with fermentation time. The scavenging activity (DPPH), total phenolic content (TPC), and total flavonoid content (TFC) of black tea kombucha were significantly higher than kombucha without tea. The porcine pancreatic lipase (PPL) inhibition of both samples increased gradually with fermentation time. Significant differences (p < 0.05) were observed in pH, TSS, and PPL inhibition from the beginning to the end of fermentation in black tea kombucha, while for kombucha without tea, significant differences (p < 0.05) were observed in pH, TFC, and PPL inhibition. Moreover, significant differences (p < 0.05) were observed in all parameters except pH between the two samples at the end of fermentation. The Pearson correlation revealed kombucha's lipase inhibition was correlated with its chemical profile, not with antioxidant activity. These findings indicate the anti-obesity potential of kombucha, highly associated with kombucha’s chemical profile.

Herbicide-resistant HPPD variants identified via directed evolution.

Herbicides play a crucial role in boosting crop yields, yet the emergence of herbicide-resistant weeds and the susceptibility of crops to herbicides have posed significant challenges to their efficacy. β-triketone herbicides specifically target the enzyme 4-Hydroxyphenylpyruvate dioxygenase (HPPD) essential for plant growth. Remarkably, few resistant weeds have been identified against these herbicides. In this study, we aimed to identify mutations within the cotton HPPD gene that confer resistance to mesotrione, a widely used triketone herbicide. Through the establishment of a high-throughput mutant screening system in E. coli, we identified four single nucleotide changes leading to amino acid substitutions in HPPD, resulting in mesotrione resistance while preserving native enzymatic activity. Various combinations of these mutations displayed synergistic effects on herbicide resistance. Additionally, the HPPD variants were able to complement the Arabidopsis athppd mutant, indicating their retention of sufficient native activity crucial for plant growth and development. Expression of these cotton HPPD variants in Arabidopsis resulted in heightened herbicide resistance. These findings offer critical insights into the target amino acids of HPPD for gene editing, paving the way for the development of herbicide-resistant cotton in the future.

Characteristics of biodepolymerized coal and insights into depolymerization pathway catalyzed by fungal extracellular enzymes

ABSTRACT The bio-depolymerization of coal is a promising technology for converting coal into low-molecular-weight industrial chemicals. In this study, we investigated the characteristics of biodepolymerized coal (BDC) and the associated depolymerization pathway. We analyzed the extracellular enzyme activities of Hypocrea lixii AH in the biodepolymerization system, performed Fourier transform infrared spectroscopy (FTIR) and 13C nuclear magnetic resonance spectroscopy (NMR) of BDC, and performed gas chromatography and mass spectrometry (GC/MS) of the BDC methanol extract. Results showed that the activities of peroxidase (POD), polyphenol oxidase (PPO), laccase (Lac), and lignin peroxidase (LiP) were 1377, 162, 10, and 1.919 U, respectively. Following biodepolymerization by Hypocrea lixii AH, 20 organic compounds were identified. Benzaldehyde 4-hydroxy-3,5-dimethoxy was the most dominant at 33.16%, followed by (Z)-9-octadecenamide, accounting for 14.73%. The aromaticity (f a) decreased from 54.3% to 23.7%, suggesting ring-opening reactions catalyzed by LiP and Lac. The ratio of carbonyl carbon (f a C) increased from 8.3% to 16.2%, suggesting oxidation reactions catalyzed by POD. By comparing the structure of the Fushun oxidized coal model and BDC, a biodepolymerization pathway was elucidated. Biodepolymerization of Fushun oxidized coal is an oxidation–depolymerization process catalyzed by a series of enzymes, which breaks the aromatic rings and oxidizes side chains. Altogether, our findings provide a theoretical support for the chemical application of BDC.

Microplastics predominantly affect gut microbiota by altering community structure rather than richness and diversity: A meta-analysis of aquatic animals

The impacts of microplastics on the gut microbiota, a crucial component of the health of aquatic animals, remain inadequately understood. This phylogenetically controlled meta-analysis aims to identify general patterns of microplastic effects on the alpha diversity (richness and Shannon index), beta diversity, and community structure of gut microbiota in aquatic animals. Data from 63 peer-reviewed articles on the Web of Science were synthesized, encompassing 424 observations across 31 aquatic species. The analysis showed that microplastics significantly altered the community structure of gut microbiota, with between-group distances being 87.75% higher than within-group distances. This effect was significant even at environmentally relevant concentrations (≤1 mg L−1). However, their effects on richness, Shannon index, and beta diversity (community variation) were found to be insignificant. The study also indicated that the effects of microplastics were primarily dependent on their concentration and size, while the phylogeny of tested species explained limited heterogeneity. Furthermore, variations in gut microbiota alpha diversity, beta diversity, and community structure were correlated with changes in antioxidant enzyme activities from the liver and hepatopancreas. This implies that gut microbiota attributes of aquatic animals may provide insights into host antioxidant levels. In summary, this study illuminates the impacts of microplastics on the gut microbiota of aquatic animals and examines the implications of these effects for host health. It emphasizes that microplastics mainly alter the community structure of gut microbiota rather than significantly affecting richness and diversity.

Comparative role of calcium oxide nanoparticles and calcium bulk fertilizer to alleviate cadmium toxicity by modulating oxidative stress, photosynthetic performance and antioxidant-defense genes expression in alfalfa

Cadmium (Cd) toxicity poses a significant threat to soil health and sustainable food production. Its bioaccumulation in plant tissues induces phytotoxicity by affecting physiological and biochemical attributes, leading to a reduction in plant biomass and production. Recently, nanotechnology has emerged as a promising approach for addressing heavy metal toxicity in an eco-friendly manner to enhance crop production. However, the comparative role of foliar applied calcium oxide nanoparticles (CaO-NPs) and bulk calcium fertilizer under Cd stress in alfalfa remains unexplored. Herein, we studied the ameliorative role of CaO-NPs and bulk calcium (50 and 100 mg L−1) to alleviate Cd stress (30 mg kg−1) in alfalfa seedlings. Plants exposed to Cd exhibited significant decreases in morpho-physiological traits, gas exchange attributes, and pigment contents as well as increase in Cd bioaccumulation in plant tissues. Notably, exogenous application of CaO-NPs ameliorates the toxic impact of Cd by enhancing plant biomass (45%), fluorescence efficiency and gaseous exchange attributes. The maximum dose of CaO-NPs induced Cd-tolerance response accompanied by a significant increase in antioxidative enzyme activities, such as superoxide dismutase (SOD; 29%), peroxidase (POD; 41%), catalase (CAT; 36%) and ascorbate peroxidase (APX; 49%), which play positive roles in ROS scavenging. TEM examination further revealed the protective role of these NPs in averting Cd-induced damage to leaf ultrastructure and mesophyll cells. Furthermore, CaO-NPs had a substantial influence on both Cd and Ca2+ accumulation in plant tissues, while qRT‒PCR analysis demonstrated higher expression of antioxidant defense genes viz. Cu/ZnSOD (0.38 fold change (FC)), MtPOD (0.51 FC), MtCAT (0.61 FC) and MtAPX (0.79 FC) under CaO-NPs application, over Cd control. Overall, our findings suggested that exogenous CaO-NPs could be effective in alleviating the adverse effects of Cd on alfalfa seedlings to ensure food safety and support sustainable agriculture.

Colorectal cancer and inflammatory bowel diseases share common salivary proteomic pathways

Inflammatory bowels diseases (IBD) are high risk conditions for colorectal cancer (CRC). The discovery of IBD and CRC noninvasive protein/peptide biomarkers using saliva and feces was the aim of this study involving 20 controls, 25 IBD (12 Crohn’s Disease-CD), 37 CRC. By untargeted proteomic (LTQ-Orbitrap/MS), a total of 152 proteins were identified in saliva. Absent in controls, 73 proteins were present in both IBD and CRC, being mainly related to cell-adhesion, cadherin-binding and enzyme activity regulation (g-Profiler). Among the remaining 79 proteins, 14 were highly expressed in CD and 11 in CRC. These proteins clustered in DNA replication/expression and innate/adaptive immunity. In stool, endogenous peptides from 30 different proteins were identified, two being salivary and CD-associated: Basic Proline-rich Protein 1 (PRBs) and Acidic Proline-rich Phosphoprotein. Biological effects of the PRBs-related peptides GQ-15 and GG-17 found in CD stool were evaluated using CRC cell lines. These peptides induced cell proliferation and activated Erk1/2, Akt and p38 pathways. In conclusion, the salivary proteome unveiled DNA stability and immunity clusters shared between IBD and CRC. Salivary PRB-derived peptides, enriched in CD stool, stimulate CRC cell proliferation and the pro-oncogenic RAS/RAF/MEK/ERK and PI3K/AKT/mTOR pathways suggesting a potential involvement of PRBs in IBD and cancer pathogenesis.

Effects of cadmium on the growth, muscle composition, digestion, gene expression of antioxidant and lipid metabolism in juvenile tilapia (Oreochromis niloticus)

Cadmium could induce various degrees of harm to aquatic organisms. A 30-day feeding trial was conducted to investigate the effects of cadmium on growth, muscle composition, digestive enzyme activity, gene expression of antioxidants and lipid metabolism in juvenile genetic improvement of farmed tilapia (GIFT, Oreochromis niloticus, Initial weight: 21.36 ± 0.24 g). Four cadmium concentrations of aquaculture water were designed: 0, 0.2, 0.4, and 0.6 mg/L Cd2+. The main results are as follows: Compared with the control group (0 mg/L Cd2+), the weight gain rate (WGR), specific growth rate (SGR), daily growth index (DGI), and spleen index (SI) of juvenile GIFT under cadmium stress were significantly decreased (p< 0.05). The contents of crude protein and crude lipid in muscle were significantly decreased (p< 0.05), and the ash was significantly increased (p< 0.05). The activities of trypsin, lipase, and α-amylase in the intestinal were significantly decreased (p< 0.05). The relative expression levels of carnitine palmityl transferase 1 (cpt-1), peroxisome proliferator-activated receptor α (pparα), pparγ, hormone-sensitive lipase (hsl), lipoprotein lipase (lpl), malate dehydrogenase (mdh), leptin (lep), fatty acid synthetase (fas), cholesterol response element binding protein 1 (srebp1), squalene cyclooxygenase (sqle), and stearoyl-CoA desaturase (scd) genes in liver were significantly decreased (p< 0.05). The relative expression levels of catalase (cat), superoxide dismutase (sod), glutathione S-transferase (gst), and glutathione peroxidase (gsh-px) genes in the liver were significantly decreased (p< 0.05). In conclusion, exposure to cadmium stress could impact growth, muscle composition, digestive enzyme activity, gene expression of antioxidant and lipid metabolism in juvenile GIFT.

Pharmacological Activities and Molecular Mechanisms of Sinapic Acid in Neurological Disorders.

Sinapic acid (SA) is a phenylpropanoid derivative found in various natural sources that exhibits remarkable versatile properties, including antioxidant, anti-inflammatory, and metal-chelating capabilities, establishing itself as a promising candidate for the prevention and treatment of conditions affecting the central nervous system, such as Alzheimer's disease (AD), Parkinson's disease (PD), ischemic stroke, and other neurological disorders. These effects also include neuroprotection in epilepsy models, as evidenced by a reduction in seizure-like behavior, cell death in specific hippocampal regions, and lowered neuroinflammatory markers. In AD, SA treatment enhances memory, reverses cognitive deficits, and attenuates astrocyte activation. SA also has positive effects on cognition by improving memory and lowering oxidative stress. This is shown by lower levels of oxidative stress markers, higher levels of antioxidant enzyme activity, and better memory retention. Additionally, in ischemic stroke and PD models, SA provides microglial protection and exerts anti-inflammatory effects. This review emphasizes SA's multifaceted neuroprotective properties and its potential role in the prevention and treatment of various brain disorders. Despite the need for further research to fully understand its mechanisms of action and clinical applicability, SA stands out as a valuable bioactive compound in the ongoing quest to combat neurodegenerative diseases and enhance the quality of life for affected individuals.

Streptomyces-Secreted Fluvirucin B6 as a Potential Bio-Fungicide for Managing Banana Fusarium Wilt and Mycotoxins and Modulating the Soil Microbial Community Structure.

Banana Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (Foc TR4) is the most destructive soil-borne fungal disease. Until now, there has been a lack of effective measures to control the disease. It is urgent to explore biocontrol agents to control Foc TR4 and the secretion of mycotoxin. In this study, fluvirucin B6 was screened from Streptomyces solisilvae using an activity-guided method. Fluvirucin B6 exhibited strong antifungal activity against Foc TR4 (0.084 mM of EC50 value) and significantly inhibited mycelial growth and spore germination. Further studies demonstrated that fluvirucin B6 could cause the functional loss of mitochondria, the disorder of metabolism of Foc TR4 cells, and the decrease of enzyme activities in the tricarboxylic acid cycle and electron transport chain, ultimately inhibiting mycotoxin metabolism. In a pot experiment, the application of fluvirucin B6 significantly decreased the incidence of banana Fusarium wilt and the amount of Foc TR4 and controlled fungal toxins in the soil. Additionally, fluvirucin B6 could positively regulate the changes in the structure of the banana rhizosphere microbial community, significantly enriching beneficial microbes associated with disease resistance. In summary, this study identifies fluvirucin B6, which plays versatile roles in managing fungal diseases and mycotoxins.

Effects of Drought Stress on Key Enzymes in Soybean Carbon and Nitrogen Metabolism

Background: The synthesis and catalysis of enzymes are inherently linked to the presence of water and drought-induced water deficit in plants can adversely impact enzyme activity. Drought severely impairs the growth and development of crops, with the extent of its impact varying according to the severity of the water scarcity. Drought also regulates the activity of carbon and nitrogen metabolizing enzymes in plants. Methods: To investigate the effects of varying degrees of drought on soybeans, this study selected HN44 (drought-resistant variety) and HN65 (drought-sensitive variety) as subjects through sand culture and assessed the changes in key enzymes involved in carbon and nitrogen metabolism within the soybean leaves in 2022. Result: The results indicated that short-term drought stress increased the activities of sucrose synthase (SUS), Sucrose Phosphate Synthase (SPS) and Glutamine Synthetase (GS), while decreasing the activity of nitrate reductase (NR). In contrast, prolonged drought reduced the activities of SUS and GS. This study provides a theoretical foundation for enhancing the drought stress resistance of soybeans.

Antioxidant Activities and Laxative Effect of Bioactive Compounds from Cynara cardunculus var. sylvestris

The present study aims to investigate the antioxidant and anti-constipation activities of the derived phenolic extracts in ribs wild cardoon (Cynara sylvestris). Analysed extracts (acetone, methanol and aqueous) exhibited high level of phenolic compounds and excellent antioxidant activities with significant IC50 values (p ≤0.05), as assessed by the DPPH radical-scavenging activity (0.84±0.08 - 2.44±0.007 μg/mL), ABTS cationic radical test (0.96 ±0.01- 136.67±6.75 μg/mL) and β-carotene bleaching assay (2.09±0.02-51.12±1.32 μg/mL). Also, higher levels of insoluble dietary fibers were found (56.18 ± 0.91% DW of neutral detergent fiber). The in vivo investigation was performed on Wistar rats to explore the ability of C. sylvestris aqueous extract (CSAE) in the enhancement of the gastrointestinal transit and the treatment of induced constipation by Loperamide (commercialized as Idium). A significant increase of 15, 24.5 and 32.4% in gastrointestinal motility was recordedwhen doses of CSAE increased (75, 150 and 300 mg/kg b.w, respectively). Food intake, water consumption, number and weight of stools were also increased in a dose-dependent manner, compared to the positive control (untreated). Moreover, CSAE provided significantly (p ≤0.05) and dose-dependently protection against oxidative stress by preserving normal antioxidant enzymes activities (SOD and GPx) in intestinal and colonic mucosa, and resorted hepatic enzymes (AST and ALT) and renal (urea and creatinine) levels to normal values. These results can be explained by the abundance of phenolic compounds and insoluble fibers in this plant. Therefore, its use can be encouraged in alimentary and pharmaceutical applications as antioxidant and laxative food supplements.

Role of Quaternary Aminoacid Derivatives and Ion Homeostasis in Amelioration of Salinity Stress

Salinity is a detriment to plant production throughout the world. Salinity impacts the quality of irrigation water. Understanding salt stress in plants necessitates complex physiological and metabolic stages. Preserving turgescent cells and regulating the water potential to maintain a proper water balance among the key responsibilities of osmoprotectants, long-term, high salt doses can slow down plant growth and development by interfering with physiological processes like nutrient imbalance, membrane damage, enzyme inhibition and metabolic dysfunctions like photosynthesis, hormone production and ion homeostasis, all of which can result in plant death. Hyperionic and hyperosmotic stress are also caused by prolonged, high salt doses. Osmolytes (such as proline and glycine betaine) accumulate under different salt levels, antioxidant enzyme activities increase and eventually some of the negative effects of NaCl toxicity on plants are mitigated. Plants under salt stress may have higher concentrations of reactive oxygen species (ROS) such as superoxide radicals (O2) and hydrogen peroxide (H2O2). Ion homeostasis is crucial for plants during abiotic stress conditions.

Coupling sensor to enzyme in the voltage sensing phosphatase

Voltage-sensing phosphatases (VSPs) dephosphorylate phosphoinositide (PIP) signaling lipids in response to membrane depolarization. VSPs possess an S4-containing voltage sensor domain (VSD), resembling that of voltage-gated cation channels, and a lipid phosphatase domain (PD). The mechanism by which voltage turns on enzyme activity is unclear. Structural analysis and modeling suggest several sites of VSD-PD interaction that could couple voltage sensing to catalysis. Voltage clamp fluorometry reveals voltage-driven rearrangements in three sites implicated earlier in enzyme activation—the VSD-PD linker, gating loop and R loop—as well as the N-terminal domain, which has not yet been explored. N-terminus mutations perturb both rearrangements in the other segments and enzyme activity. Our results provide a model for a dynamic assembly by which S4 controls the catalytic site.

Straw incorporation and nitrogen fertilization regulate soil quality, enzyme activities and maize crop productivity in dual maize cropping system

BackgroundStraw incorporation serves as an effective strategy to enhance soil fertility and soil microbial biomass carbon (SMBC), which in turn improves maize yield and agricultural sustainability. However, our understanding of nitrogen (N) fertilization and straw incorporation into soil microenvironment is still evolving. This study explored the impact of six N fertilization rates (N0, N100, N150, N200, N250, and N300) with and without straw incorporation on soil fertility, SMBC, enzyme activities, and maize yield.ResultsResults showed that both straw management and N fertilization significantly affected soil organic carbon (SOC), total N, SMBC, soil enzyme activities, and maize yield. Specifically, the N250 treatment combined with straw incorporation significantly increased SOC, total N, and SMBC compared to lower fertilization rates. Additionally, enzyme activities such as urease, cellulase, sucrose, catalase, and acid phosphatase reached their peak during the V6 growth stage in the N200 treatment under for both straw management conditions. Compared to N250 and N300 treatments of traditional planting, the N200 treatment with residue incorporation significantly increased yield by 8.30 and 4.22%, respectively. All measured parameters, except for cellulase activity, were significantly higher in spring than in the autumn across both study years, with notable increases observed in 2021.ConclusionsThese findings suggest that optimal levels of SOC, soil total N (STN), and SMBC, along with increased soil enzyme activities, is crucial for sustaining soil fertility and enhancing maize grain yield under straw incorporation and N200 treatments.

Anomalous Fluctuations in a Droplet of Chemically Active Colloids or Enzymes

Chemically active colloids or enzymes cluster into dense droplets driven by their phoretic response to collectively generated chemical gradients. Employing Brownian dynamics simulation techniques, our study of the dynamics of such a chemically active droplet uncovers a rich variety of structures and dynamical properties, including the full range of fluidlike to solidlike behavior, and non-Gaussian positional fluctuations. Our work sheds light on the complex dynamics of the active constituents of metabolic clusters, which are the main drivers of nonequilibrium activity in living systems. Published by the American Physical Society 2024

The dual role of a novel Sinorhizobium meliloti chemotaxis protein CheT in signal termination and adaptation.

Sinorhizobium meliloti senses nutrients and compounds exuded from alfalfa host roots and coordinates an excitation, termination, and adaptation pathway during chemotaxis. We investigated the role of the novel S. meliloti chemotaxis protein CheT. While CheT and the Escherichia coli phosphatase CheZ share little sequence homology, CheT is predicted to possess an α-helix with a DXXXQ phosphatase motif. Phosphorylation assays demonstrated that CheT dephosphorylates the phosphate-sink response regulator, CheY1~P by enhancing its decay two-fold but does not affect the motor response regulator CheY2~P. Isothermal Titration Calorimetry (ITC) experiments revealed that CheT binds to a phosphomimic of CheY1~P with a KD of 2.9 μM, which is 25-fold stronger than its binding to CheY1. Dissimilar chemotaxis phenotypes of the ΔcheT mutant and cheT DXXXQ phosphatase mutants led to the hypothesis that CheT exerts additional function(s). A screen for potential binding partners of CheT revealed that it forms a complex with the methyltransferase CheR. ITC experiments confirmed CheT/CheR binding with a KD of 19 μM, and a SEC-MALS analysis determined a 1:1 and 2:1 CheT/CheR complex formation. Although they did not affect each other's enzymatic activity, CheT binding to CheY1~P and CheR may serve as a link between signal termination and sensory adaptation.

Identification of Antioxidant Methyl Derivatives of Ortho-Carbonyl Hydroquinones That Reduce Caco-2 Cell Energetic Metabolism and Alpha-Glucosidase Activity

α-glucosidase, a pharmacological target for type 2 diabetes mellitus (T2DM), is present in the intestinal brush border membrane and catalyzes the hydrolysis of sugar linkages during carbohydrate digestion. Since α-glucosidase inhibitors (AGIs) modulate intestinal metabolism, they may influence oxidative stress and glycolysis inhibition, potentially addressing intestinal dysfunction associated with T2DM. Herein, we report on a study of an ortho-carbonyl substituted hydroquinone series, whose members differ only in the number and position of methyl groups on a common scaffold, on radical-scavenging activities (ORAC assay) and correlate them with some parameters obtained by density functional theory (DFT) analysis. These compounds’ effect on enzymatic activity, their molecular modeling on α-glucosidase, and their impact on the mitochondrial respiration and glycolysis of the intestinal Caco-2 cell line were evaluated. Three groups of compounds, according their effects on the Caco-2 cells metabolism, were characterized: group A (compounds 2, 3, 5, 8, 9, and 10) reduces the glycolysis, group B (compounds 1 and 6) reduces the basal mitochondrial oxygen consumption rate (OCR) and increases the extracellular acidification rate (ECAR), suggesting that it induces a metabolic remodeling toward glycolysis, and group C (compounds 4 and 7) increases the glycolysis lacking effect on OCR. Compounds 5 and 10 were more potent as α-glucosidase inhibitors (AGIs) than acarbose, a well-known AGI with clinical use. Moreover, compound 5 was an OCR/ECAR inhibitor, and compound 10 was a dual agent, increasing the proton leak-driven OCR and inhibiting the maximal electron transport flux. Additionally, menadione-induced ROS production was prevented by compound 5 in Caco-2 cells. These results reveal that slight structural variations in a hydroquinone scaffold led to diverse antioxidant capability, α-glucosidase inhibition, and the regulation of mitochondrial bioenergetics in Caco-2 cells, which may be useful in the design of new drugs for T2DM and metabolic syndrome.

A review on development of enzymatic biosensors for industrial applications

Enzymatic biosensors are vital tools across various industries due to their high specificity, sensitivity, and rapid response times. They are extensively used in medical diagnostics for monitoring biomarkers like glucose and lactate, environmental monitoring to detect pollutants, food safety and quality assurance, bioprocess control in industrial fermentation, pharmaceutical drug development and quality control, soil health and agrochemical detection in agriculture, and for identifying biological warfare agents. However, several challenges limit their broader adoption and effectiveness in industrial settings. Key challenges include enzyme stability, as enzymes can denature under extreme environmental conditions, reducing sensor lifespan and performance. Variability in enzyme activity and immobilization techniques also leads to inconsistencies, affecting sensor reproducibility and reliability. Complex sample matrices, such as those in environmental and food samples, introduce interference that can affect sensor accuracy. Integrating biosensors into existing industrial processes is another significant challenge, requiring compatibility with automated systems and real-time data processing capabilities. Additionally, high production costs and scalability issues hinder widespread adoption. Regulatory and standardization barriers further complicate the development and deployment of enzymatic biosensors. Despite these limitations, advancements in materials science, nanotechnology, and bioengineering are paving the way for more robust and reliable biosensors. Innovations such as nanomaterial-based enzyme immobilization, synthetic biology for enzyme engineering, and advanced data analytics integration hold promise for overcoming current limitations. Collaborative efforts among researchers, industry stakeholders, and regulatory bodies are essential to accelerate the development and application of enzymatic biosensors in industrial settings. While enzymatic biosensors offer significant potential, addressing these challenges through ongoing research and technological advancements is crucial for their widespread implementation and optimization.