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Over the past year, owing to the emergent demand for the search for potential COVID-19 therapeutics, identifying alternative candidates from biological sources is one of the sustainable ways to reinforce the drug discovery process. Marine macroalgae have numerous advantages because of the richest availability of underexploited bioactive compounds. Polyphenolic compounds like phlorotannins obtained from brown macroalgae are reported as proven antiviral and immunostimulatory agents. Thus, the present study evaluated the possibility of phlorotannins as antagonists to the multiple target proteins essential for SARS-CoV-2 replication. Twenty different types of potent phlorotannins were targeted against druggable target proteins, viz., 3CLpro, RdRp, and Spro using AutoDock molecular docking, drug-likeness were assessed by ADMET profiling (QikProp module). Further, validated with 200 ns molecular dynamics (MD) simulation (Desmond module) for the top-ranked phlorotannins based on docking binding affinities. Among the twenty phlorotannins studied, eckol hexacetate, phlorofucofuroeckol, fucofuroeckol, and bifuhalol-hexacetate showed significant binding affinities across the selected targets. Besides, MD simulations highlighted Glu166, Gln189, Cys145, and Thr190 tetrad as potential interaction sites to inhibit 3CLpro's activity. Moreover, phlorotannins were confirmed to be druglike, with no major deviation observed in ADMET-profiling. Hence, phlorotannins could be therapeutic candidates against SARS-CoV-2. However, further investigations are needed to prove its efficacy as an antiviral agent. Conclusively, this study may envisage that the novel finding could notably impact the advancement of antiviral interventions for COVID-19 in the near future.
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Tratamento Farmacológico da COVID-19 , SARS-CoV-2 , Humanos , Simulação de Acoplamento Molecular , Antivirais/farmacologia , Antivirais/química , Simulação de Dinâmica Molecular , Inibidores de Proteases/químicaRESUMO
Mutations in ALDH3A2 cause Sjögren-Larsson syndrome (SLS), a neuro-ichthyotic condition due to the deficiency of fatty aldehyde dehydrogenase (FALDH). We screened for novel mutations causing SLS among Indian ethnicity, characterized the identified mutations in silico and in vitro, and retrospectively evaluated their role in phenotypic heterogeneity. Interestingly, asymmetric distribution of nonclassical traits was observed in our cases. Nerve conduction studies suggested intrinsic-minus-claw hands in two siblings, a novel neurological phenotype to SLS. Genetic testing revealed five novel homozygous ALDH3A2 mutations in six cases: Case-1-NM_000382.2:c.50C>A, NP_000373.1:p.(Ser17Ter); Case-2-NM_000382.2:c.199G>T, NP_000373.1:p.(Glu67Ter); Case-3-NM_000382.2:c.1208G>A, NP_000373.1:p.(Gly403Asp); Case-4-NM_000382.2:c.1325C>T, NP_000373.1:p.(Pro442Leu); Case-5 and -6 NM_000382.2:c.1349G>A, NP_000373.1:p.(Trp450Ter). The mutations identified were predicted to be pathogenic and disrupt the functional domains of the FALDH. p.(Pro442Leu) at the C-terminal α-helix, might impair the substrate gating process. Mammalian expression studies with exon-9 mutants confirmed the profound reduction in the enzyme activity. Diminished aldehyde-oxidizing activity was observed with cases-2 and 3. Cases-2 and 3 showed epidermal hyperplasia with mild intracellular edema, spongiosis, hypergranulosis, and perivascular-interstitial lymphocytic infiltrate and a leaky eosinophilic epidermis. The presence of keratin-containing milia-like lipid vacuoles implies defective lamellar secretion with p.(Gly403Asp). This study improves our understanding of the clinical and mutational diversity in SLS, which might help to fast-track diagnostic and therapeutic interventions of this debilitating disorder.
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Aldeído Oxirredutases , Síndrome de Sjogren-Larsson , Aldeído Oxirredutases/genética , Animais , Humanos , Mamíferos/metabolismo , Mutação , Fenótipo , Estudos Retrospectivos , Síndrome de Sjogren-Larsson/genética , Síndrome de Sjogren-Larsson/patologiaRESUMO
BACKGROUND: Brown-Vialetto-Van Laere syndrome (BVVLS) and Fazio-Londe disease (FLD) are rare neurological disorders presenting with pontobulbar palsy, muscle weakness and respiratory insufficiency. Mutations in SLC52A2 (hRFVT-2) or SLC52A3 (hRFVT-3) genes can be responsible for these disorders with an autosomal recessive pattern of inheritance. The aim of this study was to screen for mutations in SLC52A2 and SLC52A3 among Indian families diagnosed with BVVLS and FLD. METHODS: SLC52A2 and SLC52A3 were screened in one FLD and three BVVLS patients by exon-specific amplification using PCR and sequencing. In silico predictions using bioinformatics tools and confocal imaging using HEK-293 cells were performed to determine the functional impact of identified mutations. RESULTS: Genetic analysis of a mother and son with BVVLS was identified with a novel homozygous mutation c.710C>T (p.Ala237Val) in SLC52A3. This variant was found to have an autosomal pseudodominant pattern of inheritance, which was neither listed in the Exome Variant Server or in the 1000 Genomes Project database. In silico analysis and confocal imaging of the p.Ala237Val variant showed higher degree of disorderness in hRFVT-3 that could affect riboflavin transport. Furthermore, a common homozygous mutation c.62A>G (p.Asn21Ser) was identified in other BVVLS and FLD patients. Despite having different clinical phenotypes, both BVVLS and FLD can be attributed to this mutation. CONCLUSION: A rare and peculiar pattern of autosomal pseudodominant inheritance is observed for the first time in two genetically related BVVLS cases with Indian origin and a common mutation c.62A>G (p.Asn21Ser) in SLC52A3 can be responsible for both BVVLS and FLD with variable phenotypes.
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Paralisia Bulbar Progressiva , Perda Auditiva Neurossensorial , Paralisia Bulbar Progressiva/genética , Células HEK293 , Perda Auditiva Neurossensorial/genética , Humanos , Proteínas de Membrana Transportadoras/genética , Mutação , Fenótipo , Receptores Acoplados a Proteínas G/genética , SíndromeRESUMO
Deficiency or defective transport of riboflavin (RF) is known to cause neurological disorders, cataract, cardiovascular anomalies, and various cancers by altering the biochemical pathways. Mechanisms and regulation of RF uptake process is well characterized in the cells of intestine, liver, kidney, and brain origin, while very little is known in the heart. Hence, we aimed to understand the expression and regulation of RF transporters (rRFVT-1 and rRFVT-2) in cardiomyocytes during RF deficiency and also investigated the role of RF in ischemic cardiomyopathy and mitochondrial dysfunction in vivo. Riboflavin uptake assay revealed that RF transport in H9C2 is (1) significantly higher at pH 7.5, (2) independent of Na+ and (3) saturable with a Km of 3.746 µM. For in vivo studies, male Wistar rats (110-130 g) were provided riboflavin deficient food containing 0.3 ± 0.05 mg/kg riboflavin for 7 weeks, which resulted in over expression of both RFVTs in mRNA and protein level. RF deprivation resulted in the accumulation of cardiac biomarkers, histopathological abnormalities, and reduced mitochondrial membrane potential which evidenced the key role of RF in the development of cardiovascular pathogenesis. Besides, adaptive regulation of RF transporters upon RF deficiency signifies that RFVTs can be considered as an effective delivery system for drugs against cardiac diseases.
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Miocárdio/metabolismo , Miócitos Cardíacos/metabolismo , Deficiência de Riboflavina/metabolismo , Riboflavina/metabolismo , Animais , Transporte Biológico Ativo , Linhagem Celular , Masculino , Proteínas de Membrana Transportadoras/metabolismo , Miocárdio/patologia , Miócitos Cardíacos/patologia , Ratos , Ratos Wistar , Deficiência de Riboflavina/patologiaRESUMO
Advanced Glycation End products (AGE) generated in a non enzymatic protein glycation process are frequently associated with diabetes, aging and other chronic diseases. Here, we explored the protective effect of phlorotannins from brown algae Padina pavonica, Sargassum polycystum and Turbinaria ornata against AGEs formation. Phlorotannins were extracted from brown algae with methanol and its purity was analyzed by TLC and RP-HPLC-DAD. Twenty five grams of P. pavonica, S. polycystum, T. ornata yielded 27.6 ± 0.8 µg/ml, 37.7 µg/ml and 37.1 ± 0.74 µg/ml of phloroglucinol equivalent of phlorotannins, respectively. Antioxidant potentials were examined through DPPH assay and their IC50 values were P. pavonica (30.12 ± 0.99 µg), S. polycystum (40.9 ± 1.2 µg) and T. ornata (22.9 ± 1.3 µg), which was comparatively lesser than the control ascorbic acid (46 ± 0.2 µg). Further, anti-AGE activity was examined in vitro by BSA-glucose assay with the extracted phlorotannins of brown algae (P. pavonica, 15.16 ± 0.26 µg/ml; S. polycystum, 35.245 ± 2.3 µg/ml; T. ornata, 22.7 ± 0.3 µg/ml), which revealed the required concentration to inhibit 50% of albumin glycation (IC50) were lower for extracts than controls (phloroglucinol, 222.33 ± 4.9 µg/ml; thiamine, 263 µg/ml). Furthermore, brown algal extracts containing phlorotannins (100 µl) exhibited protective effects against AGE formation in vivo in C. elegans with induced hyperglycemia.
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Antioxidantes/farmacologia , Caenorhabditis elegans/efeitos dos fármacos , Glucose/metabolismo , Produtos Finais de Glicação Avançada/antagonistas & inibidores , Phaeophyceae/química , Taninos/farmacologia , Animais , Antioxidantes/química , Caenorhabditis elegans/metabolismo , Produtos Finais de Glicação Avançada/análise , Produtos Finais de Glicação Avançada/metabolismo , Taninos/químicaRESUMO
Diabetes mellitus, a most common endocrine disorder of glucose metabolism, has become a global epidemic and poses a serious public health threat with an increased socio-economic burden. Escalating incidence of diabetes is correlated with changes in lifestyle and food habits that cause gut microbiome dysbiosis and ß-cells damage, which can be addressed with dietary interventions containing probiotics. Hence, the search for probiotics of human origin with anti-diabetic, anti-AGE, and anti-ACE potentials has gained renewed interest for the effective management of diabetes and its associated complications. The present study used an alloxan (AXN)-induced diabetic rat model to investigate the effects of potential probiotic Lacticaseibacillus casei MKU1, Lactiplantibacillus pentosus MKU3, and Lactiplantibacillus plantarum MKU7 administration individually on physiochemical parameters related to diabetic pathogenesis. Experimental animals were randomly allotted into six groups viz. NCG (control), DCG (AXN), DGM (metformin), DGP1 (MKU1), DGP2 (MKU3), and DGP3 (MKU7), and biochemical data like serum glucose, insulin, AngII, ACE, HbA1c, and TNF-α levels were measured until 90 days. Our results suggest that oral administration with MKU1, MKU3, or MKU7 significantly improved serum insulin levels, glycemic control, glucose tolerance, and body weight. Additionally, ß-cell mass was increased by preserving islet integrity in Lactobacillus-treated diabetic rats, whereas TNF-α (~40%), AngII (~30%), and ACE levels (~50%) were strongly inhibited and enhanced sIgA production (5.8 folds) abundantly. Furthermore, Lactobacillus administration positively influenced the gut microbiome with a significant increase in the abundance of Lactobacillus species and the beneficial Bacteroides uniformis and Bacteroides fragilis, while decreased the pathogenic Proteus vulgaris and Parabacteroides distasonis. Among the probiotic treatment groups, L. pentosus MKU3 performed greatly in almost all parameters, indicating its potential use for alleviating diabetes-associated complications.
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Microalgae, owing to their efficacy and eco-friendliness, have emerged as a promising solution for mitigating the toxicity of Bisphenol A (BPA), a hazardous environmental pollutant. This current study was focused on the degradation of BPA by Coelastrella sp. M60 at various concentrations (10-50 mg/L). Further, the metabolic profiling of Coelastrella sp. M60 was performed using GC-MS analysis, and the results were revealed that BPA exposure modulated the metabolites profile with the presence of intermediates of BPA. In addition, highest lipid (43%) and pigment content (40%) at 20 and 10 mg/L of BPA respectively exposed to Coelastrella sp. M60 was achieved and enhanced fatty acid methyl esters recovery was facilitated by Cuprous oxide nanoparticles synthesised using Spatoglossum asperum. Thus, this study persuades thepotential of Coelastrella sp. M60 for BPA degradation and suggesting new avenues to remove the emerging contaminants in polluted water bodies and targeted metabolite expression in microalgae.
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Compostos Benzidrílicos , Biodegradação Ambiental , Metabolômica , Fenóis , Compostos Benzidrílicos/metabolismo , Fenóis/metabolismo , Microalgas/metabolismo , Microalgas/efeitos dos fármacos , Cromatografia Gasosa-Espectrometria de Massas , Metaboloma , Poluentes Químicos da Água/metabolismoRESUMO
Liposomes are unique biomolecular, capable of loading both hydrophilic and hydrophobic molecules and delivered into the biological system. Liposomes (L) coated with hyaluronic acid (HA) and chitosan (CS) carrier system was fabricated. Berberine (BER) and doxorubicin (DOX) were encapsulated to enhance drug proliferation and therapeutic effect in lung cancer cells. The FTIR, XRD, SEM, and TEM techniques were carried out for functional group identification, crystallinity, and surface morphology analysis, respectively. In-vitro drug release confirms the sustained release of BER and DOX in various physiological environments. HA-CS@BER&DOX-L has good penetration ability and higher cytotoxicity effect in the A549 cells, and the IC50 value of HA-CS@BER&DOX-L is 89.19 µg/300 µL. The pure liposome showed a negligible cytotoxicity effect, and the HA-CS@BER&DOX-L could efficiently induce the apoptosis of A549 cells. The cellular uptake analysis of the HA-CS@BER&DOX-L effectively targeted and entered the A549 cells and clearly observed C. elegans images. Hence, the proposed system will be a potential treatment methodology to enhance the cytotoxicity of the A549 cancer cells and be useful to future drug administration methodology development.
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Berberina , Quitosana , Neoplasias Pulmonares , Nanopartículas , Animais , Humanos , Lipossomos/química , Neoplasias Pulmonares/tratamento farmacológico , Ácido Hialurônico/química , Berberina/farmacologia , Caenorhabditis elegans , Doxorrubicina/farmacologia , Sistemas de Liberação de Medicamentos , Nanopartículas/química , Receptores de HialuronatosRESUMO
Phycoremediation of heavy metals are gaining much attention and becoming an emerging practice for the metal removal in diverse environmental matrices. Still, the physicochemical state of metal polluted sites is often found to be complex and haphazard in nature due to the irregular discharge of wastes, that leads to the lack of conjecture on the application of microalgae for the metal bioremediation. Besides, the foresaid issues might be eventually ended up with futile effect to the polluted site. Therefore, this review is mainly focusing on interpretative assessment on pre-existing microalgal strategies and their merits and demerits for selected metal removal by microalgae through various process such as natural attenuation, nutritional amendment, chemical pretreatment, metal specific modification, immobilization and amalgamation, customization of genetic elements and integrative remediation approaches. Thus, this review provides the ideal knowledge for choosing an efficient metal remediation tactics based on the state of polluted environment. Also, this in-depth description would provide the speculative knowledge of counteractive action required for pass-over the barriers and obstacles during implementation. In addition, the most common metal removal mechanism of microalgae by adsorption was comparatively investigated with different metals through the principal component analysis by grouping various factor such as pH, temperature, initial metal concentration, adsorption capacity, removal efficiency, contact time in different microalgae. Conclusively, the suitable strategies for different heavy metals removal and addressing the complications along with their solution is comprehensively deliberated for metal removal mechanism in microalgae.
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Metais Pesados , Microalgas , Biodegradação AmbientalRESUMO
In the present study, biomass from the Chromolaena odorata plant's stem was activated using sulfuric acid to adsorb crystal violet (CV) dye. The adsorption operation of CV dye was studied considering the effect of variables like pH, initial dye concentration, time, adsorbent dosage, and temperature. The pseudo-second-order equation best fitted the kinetic study. The thermodynamic parameters such as activation energy (9.56 kJ/mol), change in Gibbs energy (81.43 to 96.7 kJ/mol), enthalpy change (6.89 kJ/mol), and entropy change (-254.4 J/mol K) were calculated. Response surface methodology estimated that at pH (4.902), adsorbent dosage (8.33 g/L), dye concentration (82.30 ppm), and temperature (300.13 K) dye removal of 97.53% is possible. FTIR, SEM, XRD, BJH, and BET confirmed adsorption operation. The adsorbent can be reused for 3 cycles effectively. Langmuir isotherm which best fitted the adsorption operation was used for designing a theoretical single-stage batch adsorber for large-scale operation.
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Chromolaena , Poluentes Químicos da Água , Violeta Genciana/química , Biomassa , Termodinâmica , Temperatura , Cinética , Adsorção , Água , Concentração de Íons de HidrogênioRESUMO
A cheap cultivation of microalgae greatly reduces the biodiesel production cost. Subsequently in this study, citric acid and effluents from sugar and tannery industries were used as the nutritional supplements for the improvement of biomass and TAG production in Phaeodactylum tricornutum using two-step cultivation. When compared to control (media without supplementation), a considerable increase in biomass and chlorophyll a was obtained with citric acid (CA) and sugar industry effluent (SIE) supplemented media. In the two-step cultivation method, biomass raised from CA (100 mg·L-1) and SIE (1.5 mL·L-1) supplementations in the first step, viz. biomass production (BP) step was allowed for lipid accumulation in the second step, viz. lipid production (LP) step, and thus yielded enhanced lipids of 11.5 ± 0.7 mg·L-1·day-1 and 13.5 ± 1.9 mg·L-1·day-1 respectively, with improved TAG synthesis. Further, differential expression analysis of TAG biosynthetic genes of P. tricornutum under single-step and two-step cultivation modes were performed, and the gene expression patterns were studied.
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Diatomáceas , Microalgas , Biocombustíveis , Biomassa , Clorofila A/metabolismo , Ácido Cítrico/metabolismo , Diatomáceas/genética , Diatomáceas/metabolismo , Lipídeos , Microalgas/genética , Microalgas/metabolismo , Açúcares/metabolismoRESUMO
Microalgae are the most attractive renewable energy sources for the production of biofuels because of their luxurious growth and lipid accumulation ability in diverse nutritional conditions. In the present study, Desmodesmus sp. VV2, an indigenous microalga, was evaluated for its biodiesel potential using Response Surface Methodology (RSM) to improve the lipid accumulation with the combination of nutrients stress NaNO3 starvation, CaCl2 depletion, and supplementation of magnesium oxide nanoparticles (MgO). Among different stress conditions, 57.6% lipid content was achieved from RSM optimized media. Owing to this, RSM results were also validated by the Artificial Neural Network (ANN) with 11 training algorithms and it is found that RSM was more significant. In addition, the saturated fatty acid (SFA) content was noticeably increased in RSM optimized media (95.8%) while compared with control. Further, the highest total FAME content 97.21% was also achieved in cells grown in RSM optimized media. Biodiesel quality parameters were analyzed and found that they are in accordance with international standards. Thus, this study suggesting that the fatty acid profile of Desmodesmus sp. VV2 attained under optimized media conditions would be suitable for biodiesel production for future energy demand.
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Biocombustíveis , Microalgas , Biomassa , Ácidos Graxos , Redes Neurais de ComputaçãoRESUMO
Marine macroalgae offer an endurable source of renewable biomass, which do not require cultivable area, fertilizers for cultivation for bioproducts production. In this study, marine brown macroalga Padina tetrastromatica as an alternate sustainable feedstock for the production of liquid fuels. Padina tetrastromatica biomass was collected from Mandapam; the coastal region of Rameswaram, Tamil Nadu, India. and the algal oil was extracted using sequential extractions using various solvents. Petroleum ether (PE) and dichloromethane (DCM) solvent fractions were found to have high lipids and further utilized for biodiesel production, wherein four different heterogeneous nanocatalysts (TiO2, Bio-Fe, GO, and MgO) and commercial homogeneous catalysts (HCl and NaOH) were employed for the transesterification. High fatty acid methyl ester (FAME) recovery (92.3%) was achieved from TiO2 mediated transesterification than the other conventional catalysts. Further, the conversion of algal biomass into bio-oil and by-products was carried out using hydrothermal liquefaction (HTL). Subsequently, the compounds were characterized by FT-IR and GC-MS analysis. The quality parameters of liquid biofuels were examined and they are in accordance with the international fuel standards. Thus, brown macroalga Padina tetrastromatica may be considered as an alternate feedstock for biofuel and other bioproducts production and TiO2 would be a suitable catalyst for the conversion of FAME.
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Biocombustíveis , Alga Marinha , Biomassa , Ácidos Graxos , Índia , Óleos de Plantas , Polifenóis , Espectroscopia de Infravermelho com Transformada de Fourier , Temperatura , ÁguaRESUMO
Novel magnetic microcomposites consisting of graphene oxide and iron oxide was synthesized to immobilize metabolically versatile Paracoccus sp. MKU1 and Leucobacter sp. AA7 and tested for the simultaneous adsorption and enhanced biological detoxification of hexavalent chromium (Cr(VI)) from tannery wastewater. This study reports highest chromium adsorption of 272.6 mg/g and 179.3 mg/g with complete reduction of Cr(VI) to Cr(III) by the microcomposites of AA7 and MKU1 from wastewater in a bioreactor (10 L) at large-scale for first time in ex situ. Furthermore, both the microcomposites displayed an enhanced detoxification of tannery wastewater by reducing various physicochemical conditions such as ammonia, nitrate, TDS, fluoride, CaCO3, Ca, Mg, NO3 and SO2 under the permissible limits. Use of electromagnetic device for magnetic microcomposites recovery from bioreactor yielded a maximum of 88% and 80.6% recovery for AA7 and MKU1, respectively. The rate of chromium recuperation achieved following desorption from the microcomposites of AA7 and MKU1 was 90.71% and 93.97%, respectively. Thus, the multifarious benefits including adsorption, metabolic detoxification, recovery, and recuperation by single functional microcomposites seems to be an intriguing and profitable approach for practicing in real-time operations to effectively remove heavy metals from the contaminated wastewater for environmental protection.
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Águas Residuárias , Poluentes Químicos da Água , Adsorção , Biodegradação Ambiental , Cromo/análise , Fenômenos Eletromagnéticos , Grafite , Fenômenos Magnéticos , Águas Residuárias/análiseRESUMO
The COVID-19 pandemic has severely destructed human life worldwide, with no suitable treatment until now. SARS-CoV-2 virus is unprecedented, resistance against number of therapeutics and spreading rapidly with high mortality, which warrants the need to discover new effective drugs to combat this situation. This current study is undertaken to explore the antiviral potential of marine algal compounds to inhibit the viral entry and its multiplication using computational analysis. Among the proven drug discovery targets of SARS-CoV-2, spike glycoprotein and 3-chymotrypsin-like protease are responsible for the virus attachment and viral genome replication in the host cell. In this study, the above-mentioned drug targets were docked with marine algal compounds (sulfated polysaccharides, polysaccharide derivatives and polyphenols) using molecular docking tools (AutoDockTools). The obtained results indicate that κ-carrageenan, laminarin, eckol, trifucol and ß-D-galactose are the top-ranking compounds showing better docking scores with SARS-CoV-2 targets, than the current experimental COVID-19 antiviral drugs like dexamethasone, remdesivir, favipiravir and MIV-150. Further, molecular dynamic simulation, ADMET and density functional theory calculations were evaluated to substantiate the findings. To the best of our knowledge, this is the first report on in silico analysis of aforesaid algal metabolites against SARS-CoV-2 targets. This study concludes that these metabolites can be curative for COVID-19 in the hour of need after further validations in in vitro and in vivo testings.Communicated by Ramaswamy H. Sarma.
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COVID-19 , SARS-CoV-2 , Humanos , Glicoproteína da Espícula de Coronavírus/farmacologia , Pandemias , Simulação de Acoplamento Molecular , Peptídeo Hidrolases , Antivirais/farmacologia , Descoberta de Drogas , Simulação de Dinâmica Molecular , Glicoproteínas , Inibidores de Proteases/farmacologiaRESUMO
Cis, cis-muconic acid (ccMA) is known for its industrial importance as a precursor for the synthesis of several biopolymers. Catechol 1,2-dioxygenase (C12O) is involved in aromatic compounds catabolism and ccMA synthesis in a greener and cleaner way. This is the first study on C12O gene from a metabolically versatile Paracoccus sp. MKU1, which was cloned and expressed in E. coli to produce ccMA from catechol. From the E. coli transformant, recombinant C12O enzyme was purified and found to be a homotrimer with a subunit size of 38.6 kDa. The apparent K m and V max for C12O was 12.89 µM and 310.1 U.mg-1, respectively, evidencing high affinity to catechol than previously reported C12Os. The predicted 3D-structure of C12O from MKU1 consisted of five α-helices in N-terminus, one α-helix in C-terminus, and nine ß-sheets in C-terminus. Moreover, a unique α-helix signature 'EESIHAN' was identified in C-terminus between 271 and 277 amino acids, however the molecular insight of conservative α-helix remains obscure. Further, fed-batch culture was employed using recombinant E. coli expressing C12O gene from Paracoccus sp. MKU1 to produce ccMA by whole-cells catalyzed bioconversion of catechol. With the successive supply of 120 mM catechol, the transformant produced 91.4 mM (12.99 g/L) of ccMA in 6 h with the purity of 95.7%. This single step conversion of catechol to ccMA using whole-cells reactions of recombinants did not generate any by-products in the reaction mixtures. Thus, the recombinant E. coli expressing high activity C12O from Paracoccus sp. MKU1 holds promise as a potential candidate for yielding high concentrations of ccMA at faster rates in low cost settings.
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Chitosan is one of the readily available polymers with relatively high abundance, biodegradable and sustainable materials with divergent functional groups that are employed in broad range of applications. Chitosan is widely used in many fields like adsorption, drug carrier for therapeutic activity, environmental remediation, drug formulation and among others. One of the unique features of chitosan is that it can be transformed to other forms like beads, films, flakes, sponges and fibres depending upon the applications. This review is aimed at showing the potential applications of chitosan and its modified solids in organic transformations. The number of existing articles is organized based on the nature of materials and subsequently with the types of reactions. After a brief description on the structural features of chitosan, properties, characterization methods including various analytical/microscopic techniques and some of the best practices to be followed in catalysis are also discussed. The next section of this review describes the catalytic activity of native chitosan without any modifications while the subsequent sections provide the catalytic activity of chitosan derivatives, chitosan covalently modified with metal complexes/salts through linkers and chitosan as support for metal nanoparticles (NPs). These sections discuss number of organic reactions that include Knoevenagel condensation, oxidation, reduction, heterocycles synthesis, cross-coupling reactions and pollutant degradation among others. A separate section provides the catalytic applications of chitosan and its modified forms for the production of fatty acid methyl esters (FAME) through esterification/transesterification reactions. The final section summarizes our views on the future directions of this field in the coming years.
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Biocombustíveis , Quitosana/química , Fulerenos/química , Catálise , Fenômenos Químicos , Técnicas de Química Sintética , Quitosana/síntese química , Complexos de Coordenação/química , Esterificação , Ácidos Graxos , Hidrólise , Nanopartículas Metálicas , Metais/química , Estrutura MolecularRESUMO
Paracoccus sp. MKU1, a metabolically versatile bacterium that encompasses diverse metabolic pathways in its genome for the degradation of aromatic compounds, was investigated for catechol bioremediation here for the first time to our knowledge. Paracoccus sp. MKU1 degraded catechol at an optimal pH of 7.5 and a temperature of 37 °C, wherein 100 mg/L catechol was completely mineralized in 96 h but required 192 h for complete mineralization of 500 mg/L catechol. While investigating the molecular mechanisms of its degradation potential, it was unveiled that Paracoccus sp. MKU1 employed both the ortho and meta pathways by inducing the expression of catechol 1,2-dioxygenase (C12O) and catechol 2,3-dioxygenase (C23O), respectively. C23O expression at transcriptional levels was significantly more abundant than C12O, which indicated that catechol degradation was primarily mediated by extradiol cleavage by MKU1. Furthermore, poly(MAA-co-BMA)-GO (PGO) microcomposites containing Paracoccus sp. MKU1 were synthesized, which degraded catechol (100 mg/L) completely within 48 h with excellent recycling performance for three cycles. Thus, PGO@Paracoccus microcomposites proved to be efficient in catechol degradation at not only faster rates but also with excellent recycling performances than free cells. These findings accomplish that Paracoccus sp. MKU1 could serve as a potential tool for bioremediation of catechol-polluted industrial wastewater and soil.
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[This corrects the article DOI: 10.1021/acsomega.8b02085.].
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BACKGROUND: Glycogen storage disease type-1a is an inherited, autosomal recessive disorder caused by mutations in G6PC1 gene leading to deficiency of glucose-6-phosphatase-α specifically in the liver/kidney/intestine. PATIENTS AND METHODS: DNA of six unrelated Indian GSD-1a patients were screened for mutations in the entire coding region of G6PC1 gene followed by direct DNA sequencing and functional was tested using glucose-6-phosphatase assay. RESULTS: Mutational screening of GSD-1a patients identified five novel mutations, viz., 1) p.V99Cfs*3, 2) p.G125R, 3) IVS1-2Aâ¯>â¯T, 4) IVS3â¯+â¯39Gâ¯>â¯A and 5) IVS3â¯+â¯42Gâ¯>â¯A along with three previously reported mutations p.G118D, p.R149Q and p.A331V. Interestingly, each of the p.V99Cfs*3, IVS1-2Aâ¯>â¯T and p.G118D mutations are identified in two unrelated GSD-1a cases. Further allelic distribution of p.V99Cfs*3 and p.A331V mutations were confirmed by RFLP analysis, consistent with autosomal recessive inheritance. Functional characterization revealed that glucose-6-phosphatase activity was completely abrogated with the mutant proteins p.G125R, p.R149Q, p.G118D, p.A331V and p.V99Cfs*3 than wild-type. However, no significant changes were observed in the expression of mutant constructs at transcription and translation level. CONCLUSION: Five novel mutations, p.V99Cfs*3, p.G125R, IVS1-2Aâ¯>â¯T, IVS3â¯+â¯39Gâ¯>â¯A and IVS3â¯+â¯42Gâ¯>â¯A are reported first time to cause GSD-1a among Indian ethnicity and are not yet reported elsewhere, suggesting separate ethnic founder effects for some mutations among Indian ethnicity.