Glucose-6-phosphate Research Articles

Evidence of a Catalytic Dyad in F420-Dependent Glucose-6-phosphate Dehydrogenase from Mycobacterium tuberculosis.

F420-dependent glucose-6-phosphate dehydrogenase (FGD) catalyzes the conversion of glucose-6-phosphate (G6P) to 6-phosphogluconolactone, using cofactor F420 as the hydride transfer acceptor. Our previous pH dependence studies suggested that E109 serves as an active site acid, donating a proton to the N-1 position of F420, while leaving the role of H40 unanswered, which was previously suggested to serve as the active site base. This work utilizes thermodynamic and kinetic studies to elucidate additional mechanistic details concerning the roles of H40 and E13. The E13 residue had not previously been considered as a key player during catalysis. Therefore, the H40A, H40Q, E13A, and E13Q FGD variants were generated and fully characterized to determine their roles in catalysis. Here, we conducted temperature-dependent pH profiles and inactivation experiments using diethylpyrocarbonate (DEPC) to determine the role of H40 during catalysis. The temperature-dependent experiments suggest that an acidic histidine can donate a proton to E13. The inactivation experiments revealed monophasic kinetics, suggesting that the one active site H40 is covalently modified by DEPC. Therefore, the active site base is a deprotonated H40 that abstracts a proton from G6P, and then a hydride is transferred to the C-5 position of cofactor F420. These data suggest that E13 and H40 act as a catalytic dyad. Global analysis of the pre-steady-state experiments revealed the accumulation of an intermediate, the spectrum of which resembles an enzyme-product complex. The global analysis also reveals fast chemistry and slow product release with cofactor association being rate-limiting in catalysis.

DEHP impairs the oxidative stress response and disrupts trace element and mineral metabolism within the mitochondria of detoxification organs.

Di(2-ethylhexyl) phthalate (DEHP), a widely utilized plasticizer in various consumer products, is classified as an endocrine disruptor and has been implicated in numerous adverse health effects, including oxidative stress, inflammation, and metabolic disturbances. Despite the growing body of literature addressing the systemic effects of DEHP, the specific influence of DEHP-induced oxidative stress on mitochondrial function within detoxification organs, particularly the liver and kidneys, remains largely unexplored. This study evaluated the effects of DEHP exposure (0, 100, 200, and 400mg/kg/day) on mitochondrial oxidative stress, trace elements, and mineral metabolism associated with signaling pathways in the liver and kidneys of rats. Altered mitochondrial oxidative stress status was indicated by impaired glucose 6-phosphate dehydrogenase (G6PD), 6-phosphoglucerate dehydrogenase (6-PGD), glutathione reductase (GR), glutathione s-transferase (GST), and glutathione peroxidase (GPx) activities, along with significant disruptions in essential minerals and trace elements, including Na, Mg, Cu, Zn, and Fe. Key oxidative stress signaling pathways, such as NF-κB, Akt, STAT3, and CREB, glucose, and tissue homeostasis, displayed dose-dependent responses to DEHP, indicating complex regulatory mechanisms. This study represents the first comprehensive investigation into DEHP-induced mitochondrial dysfunction, highlighting its effects on oxidative stress metabolism, trace element homeostasis, and cellular signaling pathways in detoxification organs. These findings provide novel insights into the mitochondrial mechanisms underlying DEHP toxicity and underscores the need for further research into the implications of plasticizer exposure on human health.

Simultaneous Observation of the Anomerization and Reaction Rates of Enzymatic Dehydrogenation of Glucose-6-Phosphate by Dissolution DNP.

The hyperpolarization of biological samples using dissolution dynamic nuclear polarization (dDNP) has become an attractive method for the monitoring of fast chemical and enzymatic reactions using NMR by taking advantage of a large signal increase. This approach is actively developing but still needs key methodological breakthroughs to be used as an analytical method for the monitoring of complex networks of simultaneous metabolic pathways. In this article, we use the deceptively simple example of glucose-6-phosphate (G6P) oxidation reaction by the enzyme G6P dehydrogenase (G6PDH) to discuss some important methodological aspects of dDNP kinetic experiments, such as its robustness and its ability to provide repeatable results as well as the capacity of this time-resolved methodology to test kinetic models and hypotheses and to provide reliable parameter estimates. To illustrate the potential of our approach, we report the first direct and quantitative evidence of selectivity of G6PDH toward the β anomer of G6P.

Mass Spectrometry Imaging Reveals Spatial Metabolic Alterations and Salidroside’s Effects in Diabetic Encephalopathy

Background: Diabetic encephalopathy (DE) is a neurological complication of diabetes marked by cognitive decline and complex metabolic disturbances. Salidroside (SAL), a natural compound with antioxidant and neuroprotective properties, has shown promise in alleviating diabetic complications. Exploring the spatial metabolic reprogramming in DE and elucidating SAL’s metabolic effects are critical for deepening our understanding of its pathogenesis and developing effective therapeutic strategies. Methods: Air-flow-assisted desorption electrospray ionization–mass spectrometry imaging (AFADESI-MSI) was employed to investigate spatial metabolic alterations in the brains of db/db mice, a spontaneous DE model. The mice were treated with SAL (30 and 150 mg/kg, orally) for 12 weeks. Differential metabolites were identified and characterized using high-resolution mass spectrometry and validated against public databases. Results: Our AFADESI-MSI analysis revealed significant changes in 26 metabolites in the brains of DE mice compared to the controls. These metabolic changes indicated disruptions in glucose, glutamate-glutamine, nucleotide, lipid, choline, aspartate, and L-carnitine metabolism. Notably, glucose 6-phosphate (G6P), glutamine, adenosine, L-carnitine, and choline exhibited similar trends in both db/db mice and STZ-induced rat models of DE, suggesting their potential as reliable biomarkers. Twelve weeks of SAL treatment demonstrated a positive regulatory effect on glucose metabolism, the glutamate–glutamine cycle, and lipid metabolism. Conclusions: This study identifies key metabolic alterations in DE and demonstrates the therapeutic potential of SAL in modulating these disturbances, offering valuable insights for targeted interventions in diabetic complications.

Synthesis, characterization, quantum chemical modelling, molecular docking, in silico and in vitro assessment of 3-(2-bromo-5-fluorophenyl))-1-(thiophen-2-yl)prop-2-en-1-one

α,β-unsaturated carbonyl compounds have extensive applications in various fields, such as organic, inorganic, analytical, and biological. In the modern era, they offer excellent pharmacological application prospects and find widespread use in the pharmaceutical industry. The current study revealed the synthesis and characterization of a novel 3-(2-bromo-5-fluorophenyl)-1-(thiophen-2-yl) prop-2-en-1-one (CY3). In vitro their antimicrobial (Pseudomonas aeruginosa, Klebsiella pneumonia, Escherichia coli, Staphylococcus aureus, and Acinetobacter baumannii), antifungal ( Candida parapsilosis, Candida tropicalis, and Candida albicans), cytotoxicity (VERO and Hep-G2 cells), in silico, and molecular docking analysis were also performed. The in-silico analysis evaluated the drug-likeness properties of the compound CY3 using various filtering rules, including Lipinski’s, Ghose filter, Veber, Egan, Muegge, and Medicinal Chemistry alerts such as Pan Assay Interference Structures (PAINS), Brenk, and Lead-likeness. Then, molecular docking studies performed using the AutoDock (AD4), Vina, and iGEMDOCK tools to determine the mechanism by which the CY3 compound interact with the bacterial strains. Here, five different receptors were selected, such as DNA gyrase, glucose 6-phosphate synthase (GlmS), dihydrofolate reductase (DHFR), dehydrosqualene synthase (DHSS), and undecaprenyl pyrophosphate synthase (UDPPS), for molecular docking analysis. The CY3 compound showed a good binding affinity with the two target proteins, DHFR and DHSS, respectively, with maximum binding energies of about − 7.07 and − 7.05 kcal/mol. The synthesized CY3 compound exhibited moderate antibacterial activity with a MIC value > 100 µg/mL against all five bacterial strains and moderate antifungal activity with a MIC value > 50 µg/mL against all three fungal strains. Drug-likeness analyses also support their favourable bioavailability.

Sulfur redirects carbon metabolism to optimize nitrogen utilization and promote andrographolide biosynthesis in Andrographis paniculata seedlings

Sulfur (S) is an important mineral nutrient element that improves plant growth and secondary metabolism. S affects the biosynthesis of andrographolide in medicinal plant Androgaphis paniculata by regulating nitrogen (N) metabolism. However, its specific role in N utilization and the connection with andrographolide biosynthesis have not yet been thoroughly understood. Here, a soilless cultivation experiment with low S (LS, 0.1 mM) and high S (HS, 2.4 mM) was conducted to investigate how S influences carbon (C) metabolism and N utilization to promote andographolide biosynthesis in Andrographis paniculata. The results showed that HS significantly increased plant biomass and N use efficiency (NUE), accompanying with remarkable enhanced andrographolide content. HS promoted the expression of photosynthetic genes, and redirected C metabolism towards to sugars accumulation by enhancing the activities of NAD-dependent glutamate dehydrogenase (NAD-GDH), malic enzyme (ME) and phosphoenolpyruvate carboxykinase (PEPCK) (P < 0.05). On the other hand, HS reduced N and S assimilation, and stimulated a greater N allocation in photosynthesis. Accordingly, NUE was increased and andrographolided biosynthesis could profit from the shift of C resource reallocation. Additionally, the significantly increased activities of ME, glucose 6-phosphate dehydrogenase (G6PDH) and isocitrate dehydrogenase (ICDH) provided reductants for secondary metabolism. HS also considerably upregulated the expression of genes in andrographolide biosynthetic pathway, including ApDXS, ApDXR, ApHDS and ApHDR in the MEP pathway, and ApGGPS. Transcription factors in the families of MYB, WRKY, ERF and bHLH, and plant hormones ABA and JA were in response to HS. Our results revealed that S synergistically promotes NUE and andrographolide biosynthesis via remodeling of C metabolism in A. paniculata.

Ferroptosis regulates hemolysis in stored murine and human red blood cells

AbstractRed blood cell (RBC) metabolism regulates hemolysis during aging in vivo and in the blood bank. However, the genetic underpinnings of RBC metabolic heterogeneity and extravascular hemolysis at population scale are incompletely understood. On the basis of the breeding of 8 founder strains with extreme genetic diversity, the Jackson Laboratory diversity outbred population can capture the impact of genetic heterogeneity in like manner to population-based studies. RBCs from 350 outbred mice, either fresh or stored for 7 days, were tested for posttransfusion recovery, as well as metabolomics and lipidomics analyses. Metabolite and lipid quantitative trait loci (QTL) mapped >400 gene-metabolite associations, which we collated into an online interactive portal. Relevant to RBC storage, we identified a QTL hotspot on chromosome 1, mapping on the region coding for the ferrireductase Steap3, a transcriptional target to p53. Steap3 regulated posttransfusion recovery, contributing to a ferroptosis-like process of lipid peroxidation, as validated via genetic manipulation in mice. Translational validation of murine findings in humans, STEAP3 polymorphisms were associated with RBC iron content, lipid peroxidation, and in vitro hemolysis in 13 091 blood donors from the Recipient Epidemiology and Donor Evaluation Study. QTL analyses in humans identified a network of gene products (fatty acid desaturases 1 and 2, epoxide hydrolase 2, lysophosphatidylcholine acetyl-transferase 3, SLC22A16, glucose 6-phosphate dehydrogenase, ELOVL, and phospholipase A2 group VI) associated with lower levels of oxylipins. These polymorphisms were prevalent in donors of African descent and were linked to allele frequency of hemolysis-linked polymorphisms for Steap3 or p53. These genetic variants were also associated with lower hemoglobin increments in thousands of single-unit transfusion recipients from the vein-to-vein database.

Stoichiometric Regeneration of Biomimetic Nicotinamide Coenzyme Powered by Biomass Sugars via In Vitro Synthetic Enzymatic Biosystems.

Biomimetic nicotinamide coenzymes, including nicotinamide mononucleotide (NMN+), have been demonstrated as promising low-cost alternatives to nicotinamide adenine dinucleotide (phosphate) (NAD(P)+) in biocatalysis. Herein, to efficiently regenerate NMNH from NMN+ in vitro powered by biomass sugars, a thermophilic NADP+-dependent glucose 6-phosphate dehydrogenase from Thermotoga maritima (TmG6PDH) was engineered to increase the activity toward NMN+. The catalytic efficiency (kcat/Km) of optimal mutant (TmG6PDH-R7) toward NMN+ increased by 71.7-fold than TmG6PDH-WT. As a result, compared to the wild type, the coenzyme specificity ([kcat/Km]NMN +/[kcat/Km]NADP +) of TmG6PDH-R7 increased by ~2.0×105-fold. The structural analysis revealed that the introduced hydrophobic and bulky residues lead to the formation of a smaller binding pocket, which resulting in a higher affinity for NMN+ with small size than NADP+. Then several in vitro synthetic enzymatic biosystems (ivSEBs) comprising this thermophilic TmG6PDH-R7 and a previously engineered thermophilic 6-phosphogluconate dehydrogenase were constructed. These ivSEBs harnessed the complete oxidation of renewable biomass sugars to facilitate the stoichiometric regeneration of 12 molecules of NMNH from 1 molecule of glucose, thereafter producing various products such as levodione, 2,3-butanediol or bioelectricity, over a wide temperature range. This study could pave the way for using stable and low-cost biomimetic coenzymes in ivSEBs for industrial biomanufacturing.

Activities of Energy and Carbohydrate Metabolism Enzymes in Rainbow Trout Оncorhynchus mykiss Walb. upon Introduction of 24-hour Lighting in Aquaculture in Southern Russia.

Activities of energy and carbohydrate metabolism enzymes (cytochrome c oxidase (COX), pyruvate kinase (PK), glucose 6-phosphate dehydrogenase (G6PD), glycerol 1-phosphate dehydrogenase (G1PDH), lactate dehydrogenase (LDH), and aldolase) were studied in rainbow trout Oncorhynchus mykiss Walb. fish grown in aquaculture in North Ossetia-Alania after introducing a regime with 24-h lighting and night feeding. COX and PK activities in the liver of fish from the experimental group were found to be significantly higher than in control fish, indicating an increase in aerobic ATP synthesis. Aldolase activity in organs of fish grown with 24-h lighting was lower than in the control fish, indicating a decrease in carbohydrate utilization in glycolysis in muscles and a lower intensity of gluconeogenesis in the liver. The differences made it possible to assume that the introduction of 24-h lighting and night feeding changed energy and carbohydrate metabolism to facilitate biosynthetic processes and, therefore, weight gain in fish.

Precision-cut liver slices as an ex vivo model to assess impaired hepatic glucose production

Fasting hypoglycemia is a severe and incompletely understood symptom of various inborn errors of metabolism (IEM). Precision-cut liver slices (PCLS) represent a promising model for studying glucose production ex vivo. This study quantified the net glucose production of human and murine PCLS in the presence of different gluconeogenic precursors. Dihydroxyacetone-supplemented slices from the fed mice yielded the highest rate, further stimulated by forskolin and dibutyryl-cAMP. Moreover, using 13C isotope tracing, we assessed the contribution of glycogenolysis and gluconeogenesis to net glucose production over time. Pharmacological inhibition of the glucose 6-phosphate transporter SLC37A4 markedly reduced net glucose production and increased lactate secretion and glycogen storage, while glucose production was completely abolished in PCLS from glycogen storage disease type Ia and Ib patients. In conclusion, this study identifies PCLS as an effective ex vivo model to study hepatic glucose production and opens opportunities for its future application in IEM research and beyond.

Optimization of a high throughput screening platform to identify inhibitors of asymmetric diadenosine polyphosphatases

When stressed, cells synthesize di-adenosine polyphosphates (ApnA), and cellular organisms also express proteins that degrade these compounds to release ATP. Most of these proteins are members of the nudix hydrolase superfamily, and several are involved in bacterial pathogenesis, neurodevelopment, and cancer. The goal of this project is to assist in the discovery of inhibitors of these enzymes that could be used to study ApnA function and the cellular role of these nudix enzymes. Because these enzymes cleave Ap4A and Ap5A to produce ATP, two standard ATP detection techniques were optimized and compared here for their suitability for high throughput screening. In the first assay, cleavage is monitored by coupling to a reaction catalyzed by firefly luciferase. In the second assay, cleavage is detected by coupling to hexokinase, glucose 6-phosphate dehydrogenase, and diaphorase. Although the former assay was more sensitive, the latter was more reproducible, linear, and suitable for screening and kinetic analyses. The assays were used to characterize the kinetics of reactions catalyzed by various nudix enzymes isolated from E. coli, humans, and Mycobacterium tuberculosis, the bacterium that causes tuberculosis. Results reveal subtle differences between the proteins that might be exploited to identify specific small molecule inhibitors.

Characterization of a Levanderina fissa Bloom in Aquaculture Ponds and Its Utilization of Dissolved Organic Phosphorus.

Harmful algal blooms (HABs) pose significant threats to ecosystems and human health worldwide, with their frequency and intensity increasing substantially. The present study reports an algal bloom observed in an aquaculture pond near Haizhou Bay in July 2022. The causative species, identified through morphological observation and DNA barcoding analysis, was the dinoflagellate Levanderina fissa (Levander) Moestrup, Hakanen, Gert Hansen, Daugbjerg & M. Ellegaard, 2014, known for causing extensive HAB events in the coastal waters of China. A sharp decline in phytoplankton species diversity was observed during the transition from the pre-bloom to the bloom phase. Furthermore, the uptake of four types of dissolved organic phosphorus (DOP), including glucose-6-phosphate (G6P), adenosine-5-triphosphate (ATP), sodium tripolyphosphate (TPP), and glyphosate, by isolated L. fissa was investigated in the laboratory. The results showed that G6P, ATP, and TPP supported L. fissa growth as effectively as orthophosphate. Additionally, the elevated concentrations of dissolved inorganic phosphorus in the media of the three treatments indicated the involvement of extracellular hydrolysis. However, alkaline phosphatase was not responsible for the hydrolysis of these three forms of DOP. This study demonstrates that the ability of L. fissa to utilize DOP may confer a competitive advantage within phytoplankton communities, potentially leading to algal blooms in aquaculture ponds.

Nephropathy induced by cisplatin results from mitochondrial disruption, impaired energy metabolism, altered expression of renal transporters, and accumulation of urinary toxins

BackgroundThe administration of platinum-based drugs such as cisplatin and its derivatives, which are frequently used during clinical chemotherapy, is highly restricted due to the incidence of nephrotoxicity. The present study focused on investigating cisplatin-induced nephrotoxicity from the perspective of energy metabolism, renal transporter expression and urinary toxin accumulation. MethodsThis study investigated cisplatin's toxic effects, including nephrotoxicity, cardiotoxicity, hepatotoxicity, pulmonary toxicity, and splenotoxicity. We used transmission electron microscopy (TEM) and scanning electron microscopy (SEM) to characterize the accumulation of cisplatin in the kidney and the structure of renal mitochondria. The production of reactive oxygen species (ROS) induced by cisplatin in renal tubular epithelial cells was evaluated by in vitro experiments, and apoptosis of renal tubular epithelial cells and alterations to the renal microvasculature were assessed. Metabolites associated with the glycolytic and tricarboxylic acid pathways were measured, and renal transporters expression, autophagy, and urinary toxins (UTs) accumulation were also assessed. ResultsOur results reveal that cisplatin-induced varying degrees of damage to the heart, liver, spleen, lungs, and kidneys, including inflammatory and fibrotic damage. Accumulation of cisplatin in renal mitochondria disrupted mitochondrial structure and mitochondrial function, as evidenced by decreased levels of glucose 6-phosphate and ribose 5-phosphate and elevated levels of isocitric acid. Cisplatin-induced accumulation of ROS in renal tubular epithelial cells led to apoptosis and, ultimately, constriction or loss of renal microvasculature. Furthermore, dysregulation of renal transporter expression, activation of autophagy and increased accumulation of UTs was observed. ConclusionAccumulation of cisplatin in the kidney led to damage to mitochondrial structure and function, apoptosis of renal tubular epithelial cells, constriction or loss of renal microvasculature, dysfunction of renal transporters, activation of autophagy, and accumulation of UTs.

Gluconeogenesis and glycogenolysis required in metastatic breast cancer cells.

Metabolic adaptability, including glucose metabolism, enables cells to survive multiple stressful environments. Glycogen may serve as a critical storage depot to provide a source of glucose during times of metabolic demand during the metastatic cascade; therefore, understanding glycogen metabolism is critical. Our goal was to determine mechanisms driving glycogen accumulation and its role in metastatic (MCF10CA1a) compared to nonmetastatic (MCF10A-ras) human breast cancer cells. 13C6-glucose flux analysis in combination with inhibitors of the gluconeogenic pathway via phosphoenolpyruvate carboxykinase (PCK), the anaplerotic enzyme pyruvate carboxylase (PC), and the rate-limiting enzyme of the pentose phosphate pathway (PPP) glucose 6-phosphate dehydrogenase (G6PD). To determine the requirement of glycogenolysis for migration or survival in extracellular matrix (ECM) detached conditions, siRNA inhibition of glycogenolysis (liver glycogen phosphorylase, PYGL) or glycophagy (lysosomal enzyme α-acid glucosidase, GAA) enzymes was utilized. Metastatic MCF10CA1a cells had 20-fold greater glycogen levels compared to non-metastatic MCF10A-ras cells. Most glucose incorporated into glycogen of the MCF10CA1a cells was in the five 13C-containing glucose (M+5) instead of the expected M+6 glycogen-derived glucose moiety, which occurs through direct glucose conversion to glycogen. Furthermore, 13C6-glucose in glycogen was quickly reduced (~50%) following removal of 13C-glucose. Incorporation of 13C6-glucose into the M+5 glucose in the glycogen stores was reduced by inhibition of PCK, with additional contributions from flux through the PPP. Further, inhibition of PC reduced total glycogen content. However, PCK inhibition increased total unlabeled glucose accumulation into glycogen, suggesting an alternative pathway to glycogen accumulation. Inhibition of the rate-limiting steps in glycogenolysis (PYGL) or glycophagy (GAA) demonstrated that both enzymes are necessary to support MCF10CA1a, but not MCF10A-ras, cell migration. GAA inhibition, but not PYGL, reduced viability of MCF10CA1a cells, but not MCF10A-ras, in ECM detached conditions. Our results indicate that increased glycogen accumulation is primarily mediated through the gluconeogenesis pathway and that glycogen utilization is required for both migration and ECM detached survival of metastatic MCF10CA1a cells. These results suggest that glycogen metabolism may play an important role in the progression of breast cancer metastasis.

Change in Activities of Enzymes of Energy and Carbohydrate Metabolism in Pink Salmon Oncorhynchus gorbuscha (Walb.) Smolts with Change in Environmental Salinity.

Activities of key enzymes of energy and carbohydrate metabolism (cytochrome c oxidase (COX), lactate dehydrogenase (LDH), aldolase, glucose 6-phosphate dehydrogenase (G6PDH), and 1-glycerophosphate dehydrogenase (1-GPDH)) were studied in pink salmon Oncorhynchus gorbuscha (Walb.) smolts from the White Sea in a cage experiment simulating the transition from a freshwater to a marine environment. A decrease in COX, G6PDH, and 1-GPDH activities and an increase in LDH and aldolase activities were observed in juveniles with an increase in water salinity. Based on the findings, a redistribution of energy substrates between the reactions of aerobic and anaerobic metabolism towards higher anaerobic ATP synthesis was assumed for pink salmon. This may indicate that adaptive mechanisms rearrange metabolism to provide energy for osmoregulation in pink salmon juveniles when the salinity changes in their habitat.

Untargeted Metabolomics and Soil Community Metagenomics Analyses Combined with Machine Learning Evaluation Uncover Geographic Differences in Ginseng from Different Locations.

Panax ginseng C.A. Meyer, known as the "King of Herbs," has been used as a nutritional supplement for both food and medicine with the functions of relieving fatigue and improving immunity for thousands of years in China. In agricultural planting, soil environments of different geographical origins lead to obvious differences in the quality of ginseng, but the potential mechanism of the differences remains unclear. In this study, 20 key differential metabolites, including ginsenoside Rb1, glucose 6-phosphate, etc., were found in ginseng from 10 locations in China using an ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS)-untargeted metabolomics approach. The soil properties were analyzed and combined with metagenomics technology to explore the possible relationships among microbial elements in planting soil. Through Spearman correlation analysis, it was found that the top 10 microbial colonies with the highest abundance in the soil were significantly correlated with key metabolites. In addition, the relationship model established by the random forest algorithm and the quantitative relationship between soil microbial abundance and ginseng metabolites were successfully predicted. The XGboost model was used to determine 20(R)-ginseng Rg2 and 2'(R)-ginseng Rg3 as feature labeled metabolites, and the optimal ginseng production area was discovered. These results prove that the accumulation of metabolites in ginseng was influenced by microorganisms in the planting soil, which led to geographical differences in ginseng quality.

Physiological and transcriptomic analyses to determine the responses of the harmful algae Akashiwo sanguinea to phosphorus utilization

Phosphorus (P) is an essential nutrient driving algal growth in aquatic ecosystems. Dissolved inorganic and organic P (DIP and DOP) are the main components in the marine P pools and are closely related to harmful algal blooms. The dinoflagellate Akashiwo sanguinea is a cosmopolitan species which frequently causes dense blooms in estuaries and coasts worldwide, while the availability of P to A. sanguinea still remain unclear. Herein, the physiological and transcriptomic responses of A. sanguinea grown under P-deficient, DIP-replete and DOP-replete conditions were compared. P-deficient adversely suppressed the growth and photosynthesis of A. sanguinea, while genes associated with P transport, DOP utilization, sulfolipid synthesis, and energy production, were markedly elevated. Three forms of DOP, namely, glucose-6-phosphate (G-6-P), adenosine 5-triphosphate (ATP), and β-Glycerol phosphate (SG-P), supported A. sanguinea growth as efficiently as DIP (NaH2PO4), and no significant difference was observed in biochemical compositions and photosynthesis of A. sanguinea between the DIP and DOP treatments. While the genes related to P transporter were markedly suppressed in DOP groups compared with the DIP group. Our results indicated that A. sanguinea is a good growth strategist under P-deficient/replete conditions, and this species had evolved a comprehensive strategy to cope with P deficiency, which might be a crucial factor driving bloom formation in a low inorganic P environment.

Evaluation of Glucose 6-Phosphate Dehydrogenase, Pyruvate Kinase, and New Generation Inflammation Biomarkers in Prolonged Neonatal Jaundice.

Background and Objectives: To evaluate the clinical findings of glucose 6-phosphate dehydrogenase (G6PD) and pyruvate kinase (PK) deficiency in prolonged jaundice and to determine whether the systemic immune inflammation index (SII), neutrophil-to-lymphocyte ratio (NLR), and platelet-to-lymphocyte ratio (PLR) can be used in the diagnosis of neonatal prolonged jaundice. Materials and Methods: Among full-term neonates with hyperbilirubinemia who were admitted to Medicine Hospital between January 2019 and January 2024 with the complaint of jaundice, 167 infants with a serum bilirubin level above 10 mg/dL, whose jaundice persisted after the 10th day, were included in this study. Results: G6PD activity was negatively correlated with NLR, SII, age, and hematocrit (Hct). There was a weak negative correlation between G6PD and NLR and a moderate negative correlation between G6PD activity and SII when adjusted for age and Hct. PK activity showed no significant correlation with G6PD, NLR, PLR, SII, age, and Hct. A linear relationship was observed between G6PD activity and SII and NLR. Conclusions: NLR and SII can be easily calculated in the evaluation of prolonged jaundice in G6PD deficiency has a considerable advantage. NLR and SII levels may contribute by preventing further tests for prolonged jaundice and regulating its treatment. It may be useful to form an opinion in emergencies and in early diagnostic period.

One-pot synthesis of γ-cyclodextrin of high purity from non-food cellulose via an in vitro ATP-free synthetic enzymatic biosystem

γ-Cyclodextrin (γ-CD) is a cyclic oligosaccharide composed of eight glucose molecules linked together via α-1,4-glycosidic bonds. It has a wide range of applications in the pharmaceutical, food, and chemical industries. Two pathways were designed for the synthesis of γ-CD from the non-food feedstock cellulose via an in vitro adenosine triphosphate (ATP) –free synthetic enzymatic biosystem. Cellulose was employed as the substrate for producing cellobiose, which was subsequently converted to γ-CD via a cascade reaction utilizing five enzymes. A stoichiometric conversion of cellulose to γ-CD was achieved by adding the synthesis module for glucose-1-phosphate (G-1-P) and optimizing the reaction conditions. The productivity of γ-CD obtained via pathway II–condition III was as high as 517 g/m3·h from cellobiose, representing a 16-fold increase compared to pathway II–condition I. A process for producing γ-CD from cellulose was established in this study, which yielded γ-CD of >90 % purity. This study presents a novel process that could be employed in next-generation biorefineries and a strategy for improving the economic considerations associated with cellulose utilization.

Dehydroepiandrosterone-α-2-Deoxyglucoside Exhibits Enhanced Anticancer Effects in MCF-7 Breast Cancer Cells and Inhibits Glucose-6-Phosphate Dehydrogenase Activity.

In the pentose phosphate pathway, dehydroepiandrosterone (DHEA) uncompetitively inhibits glucose-6-phosphate dehydrogenase (G6PD), reducing NADPH production and increasing oxidative stress, which can influence the onset and/or progression of several diseases, including cancer. 2-Deoxy-D-glucose (2-DG), a glucose mimetic, competes with glucose for cellular uptake, inhibiting glycolysis and competing with glucose-6-phosphate (G-6-P) for G6PD activity. In this study, we report that DHEA-α-2-DG (5), an α-covalent conjugate of DHEA and 2-DG, exhibits better anticancer activity than DHEA, 2-DG, DHEA +2-DG, and polydatin in MCF-7 cells, and reduces NADPH/NADP+ ratio in cellular assays. Invitro enzyme kinetics and molecular docking studies showed that 5 uncompetitively inhibits human G6PD activity and binds to the structural NADP+ site but not to the catalytic NADP+ site. Further combining 5 with the FDA-approved drug tamoxifen enhanced its cytotoxicity against MCF-7 cells, suggesting that it could serve as a candidate for combination of drug strategies.