Reductase Enzyme Research Articles

SYNTHESIS, CHARACTERIZATION, ANTIMICROBIAL AND ELECTROCHEMICAL STUDIES OF BIOSYNTHESIZED ZINC OXIDE NANOPARTICLES USING THE PROBIOTIC BACILLUS COAGULANS (ATCC 7050)

Nowadays, nanotechnology has been used to overcome many global problems such as growing worldwide demand for energy and problems of microbial antibiotic resistance. The presented study used the probiotic Bacillus coagulans (ATCC 7050) as a nano-factory for the biosynthesis of zinc oxide nanoparticles (ZnO NPs). UV-visible spectroscopy, FTIR spectroscopy, XRD, TEM and Zeta analysis confirmed the formation of spherical ZnO NPs with a mean size of 10-19 nm and positive potential of 29±2 mV. The biosynthesized ZnO NPs showed potent antimicrobial activity against Gram-positive and Gram-negative bacteria as well as pathogenic yeast with minimum inhibition concentration (MIC) values of 500 and 800 μg/ml against Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853 and 600 μg/ml against Candida albicans ATCC 10231. On the other hand, a voltaic cell composed from an immersed reduced copper (positive electrode) and platinum electrodes (negative electrode) in ZnO NPs/bacterial metabolites was connected to a voltameter and used to study the electrochemical activity of ZnO NPs/B. coagulans metabolites. Electrochemical characterization of ZnO NPs/B. coagulans metabolites was done using current density–voltage characteristic, power density and electrochemical impedance spectroscopy (EIS) analyses. ZnO NPs/B. coagulans metabolites produced high current with voltage value ≈ >0.34 volt. The present study reported the ability of B. coagulans to produce nitrate reductase enzyme with enzyme activity 2.18 U/ml. The reduction pathway of nitrate (NO3-) into nitrite (NO2-) during the biosynthesis of ZnO NPs might help and stimulate the current production.

Expression of His-tagged NADPH-dependent acetoacetyl-CoA reductase in recombinant Escherichia coli BL-21(DE3)

Poly-3-hydroxybutyrate (P(3HB)), a member of the polyhydroxyalkanoate (PHA) family, is a biodegradable polyester with diverse industrial applications. NADPH-dependent acetoacetyl-CoA reductase (phaB) is the enzyme which plays an essential role in P(3HB) synthesis by catalyzing the conversion of the intermediates. The expression of phaB enzyme using the recombinant Escherichia coli BL-21(DE3) and the purification of the synthesized enzyme were studied. The pET-B3 plasmid harbouring the phaB gene derived from Ralstonia eutropha H16, was driven by the lac promoter in E.coli BL-21(DE3). The enzyme was expressed with different induction time, temperatures and cell age. Results showed that the cell age of 4h, induction time of 12h at 37°C were identified as the optimal conditions for the enzyme reductase expression. A specific activity of 0.151 Umg-1 protein and total protein concentration of 0.518mgmg-1 of dry cell weight (DCW) were attained. Affinity chromatography was performed to purify the His-tagged phaB enzyme, in which enhanced the specific activity (14.44 Umg-1) and purification fold (38-fold), despite relative low yield (44.6%) of the enzyme was obtained. The purified phaB showed an optimal enzyme activity at 30°C and pH 8.0. The findings provide an alternative for the synthesis of the reductase enzyme which can be used in the industrial-scale production of the biodegradable polymers.

Aldehyde Dehydrogenase and Aldo-Keto Reductase Enzymes: Basic Concepts and Emerging Roles in Diabetic Retinopathy.

Diabetic retinopathy (DR) is a complication of diabetes mellitus that can lead to vision loss and blindness. It is driven by various biochemical processes and molecular mechanisms, including lipid peroxidation and disrupted aldehyde metabolism, which contributes to retinal tissue damage and the progression of the disease. The elimination and processing of aldehydes in the retina rely on the crucial role played by aldehyde dehydrogenase (ALDH) and aldo-keto reductase (AKR) enzymes. This review article investigates the impact of oxidative stress, lipid-derived aldehydes, and advanced lipoxidation end products (ALEs) on the advancement of DR. It also provides an overview of the ALDH and AKR enzymes expressed in the retina, emphasizing their growing importance in DR. Understanding the relationship between aldehyde metabolism and DR could guide innovative therapeutic strategies to protect the retina and preserve vision in diabetic patients. This review, therefore, also explores various approaches, such as gene therapy and pharmacological compounds that have the potential to augment the expression and activity of ALDH and AKR enzymes, underscoring their potential as effective treatment options for DR.

Microbial composition, rumen fermentation parameters, enteric methane emissions, and lactational performance of phenotypically high and low methane-emitting dairy cows

This experiment was designed to investigate the relation of high and low methane-yield phenotypes with body weight (BW), dry matter intake (DMI), lactation performance, enteric CH4 emissions, and rumen fermentation parameters in lactating dairy cows. A total of 130 multi- and primiparous Holstein cows were screened for enteric CH4 emissions using the GreenFeed system (C-Lock Inc.). Out of these 130 cows, 5 were identified as phenotypically high (HM) and 5 as phenotypically low (LM) CH4 emitters. Cows in the LM group had lower daily enteric CH4 emissions than cows in the HM group (on average 346 vs. 439 g/d, respectively), lower CH4 yield (15.5 vs. 20.4 g of CH4/kg of DMI), and CH4 intensity (13.2 vs. 17.0 g of CH4/ kg of energy-corrected milk yield). Enteric emissions of CO2 and H2 did not differ between HM and LM cows. These 10 cows were blocked by parity, days in milk, and milk production, and were used in a 5-wk randomized complete block design experiment. Milk composition, production, and BW were also not different between LM and HM cows. The concentration of total volatile fatty acids in ruminal contents did not differ between CH4 phenotypes, but LM cows had a lower molar proportion of acetate (57 vs. 62.1%), a higher proportion of propionate (27.5 vs. 21.6%, respectively), and therefore a lower acetate-to-propionate ratio than HM cows. Consistently, the 16S cDNA analysis revealed the abundance of Succinivibrionaceae and unclassified Veillonellaceae to be higher in LM cows compared with HM cows, bacteria that were positively correlated with ruminal propionate concentration. Notably, Succinivibrionaceae trigger the formation of propionate via oxaloacetate pathway from phosphoenolpyruvate via Enzyme Commission: 4.1.1.49, which showed a trend to be higher in LM cows compared with HM cows. Additionally, LM cows possessed fewer transcripts of a gene encoding for methyl-CoM reductase enzyme compared with HM. In this study, low and high CH4-yield cows have similar production performance and milk composition, but total-tract apparent digestibility of organic matter and fiber fractions was lower in the former group of animals.

Araucaria heterophylla oleogum resin essential oil is a novel aldose reductase and butyryl choline esterase enzymes inhibitor: in vitro and in silico evidence

The essential oil isolated by hydrodistillation of the oleogum resin of Araucaria heterophylla has been analyzed by GC–MS. Twenty-four components accounting to 99.89% of the total detected constituents of this essential oil were identified. The major ones were: caryophyllene oxide (14.8%), ( +)-sabinene (12.07%), D-limonene (11.22%), caryophyllene (10.36%), α-copaene (8.00%), β-pinene (6.44%), trans-verbenol (5.88%) and α-pinene oxide (5.18%). The in vitro inhibitory activities of this oil against aldose reductase, BuCHE, COX-2 and SARS-CoV-2 Mpro enzymes were evaluated. This revealed promising inhibitory activity of the essential oil against both aldose reductase and BuCHE enzymes. The molecular docking study of the major components of the Araucaria heterophylla essential oil was carried out to correlate their binding modes and affinities for aldose reductase and BuCHE enzymes with the in vitro results. In conclusion, the in vitro inhibitory activity of the essential oil attributed to the synergistic effect between its components and the in silico study suggested that compounds containing epoxide and hydroxyl groups may be responsible for this activity. This study is preliminary screening for the oil to be used as antidiabetic cataract and Alzheimer’s disease therapeutics and further investigations may be required.

Heterocyclic Iminoquinones and Quinones from the National Cancer Institute (NCI, USA) COMPARE Analysis.

This review uses the National Cancer Institute (NCI) COMPARE program to establish an extensive list of heterocyclic iminoquinones and quinones with similarities in differential growth inhibition patterns across the 60-cell line panel of the NCI Developmental Therapeutics Program (DTP). Many natural products and synthetic analogues are revealed as potential NAD(P)H:quinone oxidoreductase 1 (NQO1) substrates, through correlations to dipyridoimidazo[5,4-f]benzimidazoleiminoquinone (DPIQ), and as potential thioredoxin reductase (TrxR) inhibitors, through correlations to benzo[1,2,4]triazin-7-ones and pleurotin. The strong correlation to NQO1 infers the enzyme has a major influence on the amount of the active compound with benzo[e]perimidines, phenoxazinones, benz[f]pyrido[1,2-a]indole-6,11-quinones, seriniquinones, kalasinamide, indolequinones, and furano[2,3-b]naphthoquinones, hypothesised as prodrugs. Compounds with very strong correlations to known TrxR inhibitors had inverse correlations to the expression of both reductase enzymes, NQO1 and TrxR, including naphtho[2,3-b][1,4]oxazepane-6,11-diones, benzo[a]carbazole-1,4-diones, pyranonaphthoquinones (including kalafungin, nanaomycin A, and analogues of griseusin A), and discorhabdin C. Quinoline-5,8-dione scaffolds based on streptonigrin and lavendamycin can correlate to either reductase. Inhibitors of TrxR are not necessarily (imino)quinones, e.g., parthenolides, while oxidising moieties are essential for correlations to NQO1, as with the mitosenes. Herein, an overview of synthetic methods and biological activity of each family of heterocyclic imino(quinone) is provided.

Efficient Dye Degradation and Antibacterial Activity of Carbon Dots/Chitosan-Doped La2O3 Nanorods: In Silico Molecular Docking Analysis.

This work demonstrates the degradation of toxic RhB (rhodamine B) dye from polluted water in various pH environments. It assesses the antibacterial action of CDs (carbon dots)/CS (chitosan)-doped La2O3 (lanthanum oxide) NRs (nanorods). CS and CDs have been introduced as dopants to modify the characteristics of La2O3 to achieve efficient outcomes. The influence of doping on the structural, morphological, optical, and elemental properties of synthesized La2O3 NRs was investigated through a number of analytical techniques. The structural analysis of XRD revealed a hexagonal phase. The rod-like structure of pure La2O3 and reduction in the size of NRs upon doping were exhibited by TEM micrographs. From UV-vis spectroscopy, increased absorption upon doping and introduction of redshift that led to reduced bandgap energy were observed. The FTIR spectra indicate the presence of functional groups of pure and integrated samples. The catalytic activity of specimens in basic medium toward dye showed excellent results (94.57%). The inhibition zone of diameter 4.15 mm was evaluated by 6 mL of CDs/CS-doped La2O3 NRs against Escherichia coli once the surface area increased by dopants. In silico experiments were performed for enoyl-[acyl-carrier-protein] reductase (FabI) and DNA gyrase enzymes to assess the potency of CS-doped La2O3 and CDs/CS-doped La2O3 as their inhibitors and to justify their possible mechanism of action.

Computational Analysis to Predict Drug Targets for the Therapeutic Management of Mycobacterium avium sub. Paratuberculosis.

Mycobacterium avium sp. paratuberculosis (MAP) is a pathogen, which causes paratuberculosis in animals; it has also been found to be associated with a number of autoimmune disorders in humans. The emergence of drug resistance has also been found in this bacillus during disease management. The present study's focus was to identify potential therapeutic targets for the therapeutic management of Mycobacterium avium sp. paratuberculosis infection by in silico analysis. Differentially-expressed genes (DEGs) can be good drug targets, which can be identified from microarray studies. We used gene expression profile GSE43645 to identify differentiallyexpressed genes. An integrated network of upregulated DEGs was constructed with the STRING database and the constructed network was analyzed and visualized by Cytoscape. Clusters in the proteinprotein interaction (PPI) network were identified by the Cytoscape app ClusterViz. MAP proteins predicted in clusters were analyzed for their non-homology with the human proteins, and homologous proteins were excluded. Essential proteins and cellular localization analysis and the physicochemical characteristics prediction were also done. Finally, the druggability of the target proteins and drugs that can block the targets was predicted using the DrugBank database and confirmed by molecular docking. Structural prediction and verification of drug target proteins were also carried out. Two drug targets, MAP_1210 (inhA) and MAP_3961 (aceA), encoding enoyl acyl carrier protein reductase and isocitrate lyase enzymes, respectively, were finally predicted as potential drug targets. Both of these proteins have been predicted as drug targets in other mycobacterial species also, supporting our results. However, further experiments are required to confirm these results.

Efektivitas Ekstrak Daun Annona Muricata Terhadap Penurunan Kolesterol Total Pada Lansia Dengan Hiperkolesterolemia Di Panti Werdha Mojopahit Mojokerto

The elderly experience a decrease in function, especially in the function of blood circulation. Blood circulation diseases and hormonal metabolism disorders experienced by the elderly include hypercholesterolemia. Hypercholesterolemia is a disorder of fat metabolism that triggers an increase in total blood cholesterol levels, namely> 200 mg/dl. The study aimed to determine the effectiveness of A. muricata leaf extract in decreasing total cholesterol levels in the elderly with hypercholesterolemia. Quantitative research method, using a quasi-experimental type with a group pretest-posttest design, with a sample size of 20 elderly. The sampling technique used purposive sampling, was carried out for a week consuming three times a day. The paired t-test showed a value of 8.73 and a p-value of 0.001. This study's results indicated a significant difference between the average cholesterol levels in the pre-test and post-test administration of Annona Muricata leaf extract. Compounds contained in soursop leaves, namely flavonoids, by reducing the activity of HMG-koa reductase (enzyme β-Hydroxy β-methylglutaryl-CoA) can reduce the activity of ACAT (enzyme acyl-Coa cholesterol acyltransferase), and reduce the absorption activity of cholesterol in the liver. Digestive tract. Saponin compounds in soursop leave lower cholesterol by binding to cholesterol in the intestinal lumen, thereby preventing the reabsorption of cholesterol. Saponins can bind with bile acids, which in turn will reduce the enterohepatic circulation of bile acids and cause an increase in the process of excretion of cholesterol.

Prdx5 in the Regulation of Tuberous Sclerosis Complex Mutation-Induced Signaling Mechanisms.

(1) Background: Tuberous sclerosis complex (TSC) mutations directly affect mTORC activity and, as a result, protein synthesis. In several cancer types, TSC mutation is part of the driver mutation panel. TSC mutations have been associated with mitochondrial dysfunction, tolerance to reactive oxygen species due to increased thioredoxin reductase (TrxR) enzyme activity, tolerance to endoplasmic reticulum (ER) stress, and apoptosis. The FDA-approved drug rapamycin is frequently used in clinical applications to inhibit protein synthesis in cancers. Recently, TrxR inhibitor auranofin has also been involved in clinical trials to investigate the anticancer efficacy of the combination treatment with rapamycin. We aimed to investigate the molecular background of the efficacy of such drug combinations in treating neoplasia modulated by TSC mutations. (2) Methods: TSC2 mutant and TSC2 wild-type (WT) cell lines were exposed to rapamycin and auranofin in either mono- or combination treatment. Mitochondrial membrane potential, TrxR enzyme activity, stress protein array, mRNA and protein levels were investigated via cell proliferation assay, electron microscopy, etc. (3) Results: Auranofin and rapamycin normalized mitochondrial membrane potential and reduced proliferation capacity of TSC2 mutant cells. Database analysis identified peroxiredoxin 5 (Prdx5) as the joint target of auranofin and rapamycin. The auranofin and the combination of the two drugs reduced Prdx5 levels. The combination treatment increased the expression of heat shock protein 70, a cellular ER stress marker. (4) Conclusions: After extensive analyses, Prdx5 was identified as a shared target of the two drugs. The decreased Prdx5 protein level and the inhibition of both TrxR and mTOR by rapamycin and auranofin in the combination treatment made ER stress-induced cell death possible in TSC2 mutant cells.

Potential Role of Ribonucleotide Reductase Enzyme in Mitochondria Function and Woody Breast Condition in Broiler Chickens.

The cellular events leading to the development of the woody breast myopathy in broiler breast muscle are unclear. Affected woody breast muscle exhibits muscle fiber degeneration/regeneration, connective tissue accumulation, and adverse morphological changes in mitochondria. Ribonucleotide reductase (RNR) is an enzyme for the synthesis of dNTP, which is important for mitochondria DNA content (mtDNA). RNR consists of two subunits: RRM1/RRM2. A decrease in RRM2 is associated with a decrease in mtDNA and mitochondria proteins, leading to impaired ATP production. The objective of this study was to investigate potential RNR differences between woody breast (WB) and normal (N) breast muscle by examining RRM2 expression and associated pathways. Gene expression and enzyme activities were examined by qPCR and commercial kits. Results showed that RRM2 expression reduced for WB (p = 0.01) and genes related to mitochondria, including ATP6 (p = 0.03), COX1 (p = 0.001), CYTB (p = 0.07), ND2 (p = 0.001) and ND4L (p = 0.03). Furthermore, NDUFB7 and COX 14, which are related to mitochondria and ATP synthesis, tended to be reduced in WB. Compared to N, GLUT1 reduced for WB (p = 0.05), which is responsible for glucose transport in cells. Consequently, PDK4 (p = 0.0001) and PPARG (p = 0.008) increased in WB, suggesting increased fatty acid oxidation. Citric synthase activity and the NAD/NADH ratio (p = 0.02) both reduced for WB, while WB increased CHRND expression (p = 0.001), which is a possible indicator of high reactive oxygen species levels. In conclusion, a reduction in RRM2 impaired mitochondria function, potentially ATP synthesis in WB, by increasing fibrosis and the down-regulation of several genes related to mitochondria function.

Synthesis, antidiabetic activity and molecular docking studies of novel aryl benzylidenethiazolidine-2,4-dione based 1,2,3-triazoles.

A series of novel aryl benzylidenethiazolidine-2,4-dione based 1,2,3-triazoles synthesized in a straightforward route consisting of benzylidenethiazolidine-2,4-dione and 1,2,3-triazole pharmacophores. The new scaffolds tested for in vitro antidiabetic activity by inhibition of aldose reductase enzyme and its inhibition measured in half of Inhibition Concentration (IC50). The activity results correlated with standard reference Sorbinil (IC50: 3.45 ± 0.25µM). Among all the titled compounds 8f (1.42 ± 0.21µM), 8d (1.85 ± 0.39µM), 13a (1.94 ± 0.27µM) and 8b (1.98 ± 0.58µM) shown potent activity. In addition, molecular docking results against the crystal structure of aldose reductase (PDB ID: 1PWM) revealed that the binding affinities shown by all synthesized compounds are higher than the reference compound Sorbinil. The docking scores, H-bond interactions, and hydrophobic interactions well defined inhibition strength of all compounds.

A Review on Hepatotoxic Effects of Methotrexate Therapy with Possible Strategies that May Counteract Hepatotoxicity

Methotrexate, inhibitor of a dihydrofolate reductase, is a chemotherapeutic treatment applied in many kinds of tumors as an anti-metabolite. also, Methotrexate applied to treat different other disorders, including diseases of autoimmunity, such as psoriasis, rheumatism, vasculitis and ulcerative colitis and Crohn’s disease. However, Methotrexate induces toxic effects on neurons, kidney toxicity, and liver damage, also Methotrexate have been associated with elevation risk of hepatic injury, fibrosis and cirrhosis. Methotrexate work as a dihydrofolic acid analog that binds to the dihydrofolate reductase enzyme that prevent the synthesis of tetrahydrofolic acid, that is essential synthesis of DNA. Although folic acid combine with Methotrexate therapy to prevent hepatotoxic effects, folic acid may reduce therapeutic effects of Methotrexate so recent research focus incorporation of new agent to counteract Methotrexate induced hepatotoxicity that include vitamin B12, vitamin E, N-acetylcysteine, melatonin, Erdosteine, inulin, Ferulic acid, Human placental extract, Natural plant extract (flavonoids). These agents are promising to counteract Methotrexate induced hepatotoxicity. Materials and methods: A comprehensive literature review on different published articles hepatotoxic effects of methotrexate and was carried out by electronic research on Medline, PubMed, Google scholar, and Cochrane Libran data bases using different searching terms related to prevention of methotrexate induced hepatotoxicity. The article focusses on clinical studies in different regions in the period of 2018-2022. Objective: To search for new update modality preventive therapy for methotrexate induced liver damage Results: Recent study oriented for the prevention and treatment of methotrexate associated with hepatotoxicity.

The therapeutic effect of wasp venom (Vespa magnifica, Smith) and its effective part on rheumatoid arthritis fibroblast-like synoviocytes through modulating inflammation, redox homeostasis and ferroptosis

Ethnopharmacological relevanceRheumatoid arthritis (RA) is a chronic inflammatory disease that is related to the aberrant proliferation of fibroblast-like synoviocytes (FLS). Wasp venom (WV, Vespa magnifica, Smith), an insect secretion, has been used to treat RA in Chinese Jingpo national minority's ancient prescription. However, the potential mechanisms haven't been clarified. Aim of the studyThe purposes of this paper were two-fold. First, to investigate which was the best anti-RA effective part of WV-I (molecular weight less than 3 kDa), WV-II (molecular weight 3–10 kDa) and WV-III (molecular weight more than 10 kDa) that were separated from WV. Second, to explore the underlying molecular mechanism of WV and WV-II that was best effective part in RA. Materials and methodsThe wasps were electrically stimulated and the secretions were collected. WV-I, WV-II and WV-III were acquired by ultracentrifuge method according to molecular weight. Next, WV, WV-I, WV-II and WV-III were identified by HPLC. Functional annotation and pathway analysis of WV used to bioinformatics analysis. RNA-seq analyses were constructed to identify differentially expressed genes (DEGs). GO and KEGG pathway analyses were performed by Metascape database. STRING was used to analyze the PPI network from DEGs. Next, PPI network was visualized using Cytoscape that based on MCODE. The pivotal genes of PPI network and MCODE analysis were verified by qRT-PCR. Subsequently, MH7A cells were performed by MTT assay to evaluate the ability of inhibiting cell proliferation. Luciferase activity assay was conducted in HepG2/STAT1 or HepG2/STAT3 cells to assess STAT1/3 sensitivity of WV, WV-I, WV-II and WV-III. Additionally, interleukin (IL)-1β and IL-6 expression levels were detected by ELISA kits. Intracellular thioredoxin reductase (TrxR) enzyme was evaluated by TrxR activity assay kit. ROS levels, lipid ROS levels and Mitochondrial membrane potential (MMP) were assessed by fluorescence probe. Cell apoptosis and MMP were measured by using flow cytometry. Furthermore, the key proteins of JAK/STAT signaling pathway, protein levels of TrxR and glutathione peroxidase 4 axis (GPX4) were examined by Western blotting assay. ResultsRNA-sequencing analysis of WV displayed be related to oxidation-reduction, inflammation and apoptosis. The data displayed that WV, WV-II and WV-III inhibited significantly cells proliferation in human MH7A cell line compared to WV-I treatment group, but WV-III had no significant suppressive effect on luciferase activity of STAT3 compared with IL-6-induced group. Combined with earlier reports that WV-III contained major allergens, we selected WV and WV-II further to study the mechanism of anti-RA. In addition, WV and WV-II decreased the level of IL-1β and IL-6 in TNF-α-induced MH7A cells via inactivating of JAK/STAT signaling pathway. On the other hand, WV and WV-II down-regulated the TrxR activity to produce ROS and induce cell apoptosis. Furthermore, WV and WV-II could accumulate lipid ROS to induce GPX4-mediated ferroptosis. ConclusionsTaken together, the experimental results revealed that WV and WV-II were potential therapeutic agents for RA through modulating JAK/STAT signaling pathways, redox homeostasis and ferroptosis in MH7A cells. Of note, WV-II was an effective part and the predominant active monomer in WV-II will be further explored in the future.

3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitory activity of Indonesian Cajanus cajan leaves and Zingiber officinale extracts

One indicator of antihypercholesterolemic candidates is their inhibitory activity on 3hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase enzyme. This study aimed to analyze the HMG-CoA reductase inhibitory activities of Cajanus cajan leaves, Zingiber officinale var Amarum and Zingiber officinale var Rubrum. C. cajan leaves and two types of Z. officinale were extracted using the maceration method with three solvents: distilled water, 70% ethanol and 96% ethanol. All nine types of resulted extracts were then analyzed for bioactive compounds and tested for the HMG-CoA reductase inhibitory activity using the HMG-CoA reductase analysis kit (BioVision’s K588-100). It was carried out in vitro using a colorimetric microplate reader with a wavelength of 340 nm at 37°C. The results showed that the extracts contained flavonoids, tannins, saponins and steroids. The nine extracts showed inhibitory activities on the HMG-CoA reductase enzyme. The three extracts showed the highest inhibitory activity; 70% ethanolic Z. officinale var Amarum extract (47.87%), aqueous Z. officinale var Rubrum extract (40.62%) and 70% ethanolic C. cajan leaves extract (39.38%). These results indicate that the three extracts have a medium inhibitory level of HMG-CoA reductase. The study suggested that the three types of extracts could be used as antihypercholesterolemic candidates such as antihypercholesterolemic functional food.

Recoding UAG to selenocysteine in Saccharomyces cerevisiae.

Unique chemical and physical properties are introduced by inserting selenocysteine (Sec) at specific sites within proteins. Recombinant and facile production of eukaryotic selenoproteins would benefit from a yeast expression system; however, the selenoprotein biosynthetic pathway was lost in the evolution of the kingdom Fungi as it diverged from its eukaryotic relatives. Based on our previous development of efficient selenoprotein production in bacteria, we designed a novel Sec biosynthesis pathway in Saccharomyces cerevisiae using Aeromonas salmonicida translation components. S. cerevisiae tRNASer was mutated to resemble A. salmonicida tRNASec to allow recognition by S. cerevisiae seryl-tRNA synthetase as well as A. salmonicida selenocysteine synthase (SelA) and selenophosphate synthetase (SelD). Expression of these Sec pathway components was then combined with metabolic engineering of yeast to enable the production of active methionine sulfate reductase enzyme containing genetically encoded Sec. Our report is the first demonstration that yeast is capable of selenoprotein production by site-specific incorporation of Sec.

Assessment of the Anti-Breast Cancer Effects of Urolithin with Molecular Docking Studies in the In Vitro Condition: Introducing a Novel Chemotherapeutic Drug.

A lot of research has been done on using natural items as diabetes treatment. The molecular docking study was conducted to evaluate the inhibitory activities of urolithin A against α-amylase, α-glucosidase, and aldose reductase. The molecular docking calculations indicated the probable interactions and the characteristics of these contacts at an atomic level. The results of the docking calculations showed the docking score of urolithin A against α-amylase was -5.169kcal/mol. This value for α-glucosidase and aldose reductase was -3.657kcal/mol and -7.635kcal/mol, respectively. In general, the outcomes of the docking calculations revealed that urolithin A can construct several hydrogen bonds and hydrophobic contacts with the assessed enzymes and reduces their activities considerably. The properties of urolithin against common human breast cancer cell lines, i.e., SkBr3, MDA-MB-231, MCF-7, Hs578T, Evsa-T, BT-549, AU565 and 600MPE were evaluated. The IC50 of the urolithin was 400, 443, 392, 418, 397, 530, 566 and 551 against SkBr3, MDA-MB-231, MCF-7, Hs578T, Evsa-T, BT-549, AU565 and 600MPE, respectively. After doing the clinical trial studies, the recent molecule may be used as an anti-breast cancer supplement in humans. IC50 values of urolithin A on α-amylase, α-glucosidase, and aldose reductase enzymes were obtained at 16.14, 1.06 and 98.73µM, respectively.

In vitro and in silico antioxydant activity, toxicity prediction, and molecular docking study of 3- and 3,3'-nitrophenylferrocene and their reduced amines

The antioxidant activity of 3-nitrophenylferrocene (3NPF) and 3,3'-nitrophenylferrocene (3,3'NPF) and their reduced amines was measured using superoxide anion radical (). Binding parameters such as binding free energies and binding constants were also calculated. sign and values suggest respectively the spontaneity and a strong interaction between the radical and all studied compounds. Molecular docking study showed that 3NPF is most inreactive compound against glutathione reductase enzyme having the the lowest docking scores of -16.96 kJ/mol. The two reduced forms were predected to be non-toxic and are not inhibitors of CYP450 2C19, 2D6 isoenzymes which suggests a decrease in their plasma concentrations and a rapid elimination route.

Pengukuran Aktivitas Nitrat Reduktase (ANR) Pada Tanaman Poaceae Secara In vivo

Corn, rice, and sugarcane are included in the Poaceae family which have high economic value as staple foods. Nitrate reductase plays a role in reducing nitrate to ammonia. The nitrate reductase activity testing can help biotechnology studies, in this case, fertilizer production or organic waste treatment, because the nitrate reductase enzyme can be used to convert nitrate into more useful nitrogen compounds. The purpose of this study was to analyze the activity of nitrate reductase (ANR) from the leaves of the Poaceae family, namely corn, rice, and sugarcane. In vivo ANR measurements were carried out using the spectrophotometric method with two types of spectrophotometers, namely the Vis/UV Spectrophotometer, Genesys10UV, and Visible Spectrophotometer, with a wavelength of 540 nm. This study concludes that rice had the highest ANR of 0.1503 µmol/gram leaf weight/hour of incubation. The increase in nitrate reductase activity indicates that the energy for nitrate reduction also increases and provides a large capacity for synthesizing amino acids, proteins, or total N assimilation.

Inhibitory effect of herbal compounds on the Oxygen-insensitive NADPH nitro reductase enzyme of metronidazole-resistant Helicobacter pylori

Helicobacter pylori is a significant risk factor for chronic gastritis, gastric ulcers, and gastric cancer. The purpose of this article is to investigate the potential impact of fifty herbal compounds derived from Ginger and Parsley plants, known for their antibacterial properties on the Oxygen-insensitive NADPH nitro reductase enzyme of metronidazole-resistant H. pylori. In the present study, the information on the structure of compounds, the H. pylori resistant to metronidazole enzyme, myristicin, and shogaol derivatives were obtained from databases such as ZINC15, RCSB (Protein Data Bank), and PubChem, respectively. Finally, molecular docking was performed with iGemdock2.1 and Molegro Virtual Docker. After molecular docking, four out of the fifty phytocompounds showed the lowest energy and the highest number of interactions with the amino acids at the binding sites. Among these four phytocompounds, the best phytocompound was N-Vanillyloctanamide derived from Ginger. Our molecular docking study suggests that ginger can be introduced as a potential candidate to inhibit the growth of H. pylori.