RESUMO
Microbial epoxide hydrolases, cis-epoxysuccinate hydrolases (CESHs), have been utilized for commercial production of enantiomerically pure L(+)- and D(-)-tartaric acids for decades. However, the stereo-catalytic mechanism of CESH producing L(+)-tartaric acid (CESH[L]) remains unclear. Herein, the crystal structures of two CESH[L]s in ligand-free, product-complexed, and catalytic intermediate forms were determined. These structures revealed the unique specific binding mode for the mirror-symmetric substrate, an active catalytic triad consisting of Asp-His-Glu, and an arginine providing a proton to the oxirane oxygen to facilitate the epoxide ring-opening reaction, which has been pursued for decades. These results provide the structural basis for the rational engineering of these industrial biocatalysts.
Assuntos
Biocatálise , Epóxido Hidrolases , Hidrolases , Epóxido Hidrolases/metabolismo , Hidrolases/química , Hidrolases/genética , Hidrolases/metabolismo , Tartaratos/metabolismo , Modelos Moleculares , Estrutura Terciária de Proteína , Estrutura Quaternária de ProteínaRESUMO
l-(+)-Tartaric acid plays important roles in various industries, including pharmaceuticals, foods, and chemicals. cis-Epoxysuccinate hydrolases (CESHs) are crucial for converting cis-epoxysuccinate to l-(+)-tartrate in the industrial production process. There is, however, a lack of detailed structural and mechanistic information on CESHs, limiting the discovery and engineering of these industrially relevant enzymes. In this study, we report the crystal structures of RoCESH and KoCESH-l-(+)-tartrate complex. These structures reveal the key amino acids of the active pocket and the catalytic triad residues and elucidate a dynamic catalytic process involving conformational changes of the active site. Leveraging the structural insights, we identified a robust BmCESH (550 ± 20 U·mg-1) with sustained catalytic activity even at a 3 M substrate concentration. After six batches of transformation, immobilized cells with overexpressed BmCESH maintained 69% of their initial activity, affording an overall productivity of 200 g/L/h. These results provide valuable insights into the development of high-efficiency CESHs and the optimization of biotransformation processes for industrial uses.
Assuntos
Biocatálise , Tartaratos , Tartaratos/metabolismo , Tartaratos/química , Domínio Catalítico , Cristalografia por Raios X , Hidrolases/química , Hidrolases/metabolismo , Hidrolases/genética , Modelos Moleculares , Conformação ProteicaRESUMO
In rice paddies, soil and plant-derived organic matter are degraded anaerobically to methane (CH4), a powerful greenhouse gas. The highest rate of methane emission occurs during the reproductive stage of the plant when mostly dicarboxylic acids are exudated by the roots. The emission of methane at this stage depends largely on the cooperative interaction between dicarboxylic acid-fermenting bacteria and methanogenic archaea in the rhizosphere. The fermentation of tartrate, one of the major acids exudated, has been scarcely explored in rice paddy soils. In this work, we characterized an anaerobic consortium from rice paddy soil composed of four bacterial strains, whose principal member (LT8) can ferment tartrate, producing H2 and acetate. Tartrate fermentation was accelerated by co-inoculation with a hydrogenotrophic methanogen. The assembled genome of LT8 possesses a Na+-dependent oxaloacetate decarboxylase and shows that this bacterium likely invests part of the H2 produced to reduce NAD(P)+ to assimilate C from tartrate. The phylogenetic analysis of the 16S rRNA gene, the genome-based classification as well as the average amino acid identity (AAI) indicated that LT8 belongs to a new genus within the Sporomusaceae family. LT8 shares a few common features with its closest relatives, for which tartrate degradation has not been described. LT8 is limited to a few environments but is more common in rice paddy soils, where it might contribute to methane emissions from root exudates.IMPORTANCEThis is the first report of the metabolic characterization of a new anaerobic bacterium able to degrade tartrate, a compound frequently associated with plants, but rare as a microbial metabolite. Tartrate fermentation by this bacterium can be coupled to methanogenesis in the rice rhizosphere where tartrate is mainly produced at the reproductive stage of the plant, when the maximum methane rate emission occurs. The interaction between secondary fermentative bacteria, such as LT8, and methanogens could represent a fundamental step in exploring mitigation strategies for methane emissions from rice fields. Possible strategies could include controlling the activity of these secondary fermentative bacteria or selecting plants whose exudates are more difficult to ferment.
Assuntos
Euryarchaeota , Oryza , Solo/química , Oryza/microbiologia , Fermentação , Tartaratos/metabolismo , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo , Filogenia , Composição de Bases , Análise de Sequência de DNA , Bactérias , Bactérias Anaeróbias/metabolismo , Euryarchaeota/metabolismo , Firmicutes/metabolismo , Bactérias Gram-Negativas/genética , Metano/metabolismoRESUMO
OBJECTIVE: Xenobiotic metabolites are exogenous biochemicals that can adversely impact reproductive health. We previously identified xenobiotics in cervicovaginal fluid during pregnancy in association with short cervix. In other organ systems, xenobiotics can modify epithelial barrier function. We hypothesise that xenobiotics dysregulate epithelial cell and macrophage immune responses as a mechanism to disrupt the cervicovaginal barrier. DESIGN: In vitro cell culture system. SETTING: Laboratory within academic institution. SAMPLE: Vaginal, ectocervical and endocervical epithelial cell lines and primary macrophages. METHODS: Cells were treated with diethanolamine (2.5 mM), ethyl glucoside (5 mM) or tartrate (2.5 mM) for 24 h. MAIN OUTCOME MEASURES: Cytokines and matrix metalloproteinases were measured in cell supernatants (n = 3 per condition). One-way analysis of variance (ANOVA) with Dunnett's test for multiple comparisons was performed. RESULTS: Diethanolamine induces inflammatory cytokines, whereas ethyl glucoside and tartrate generally exert anti-inflammatory effects across all cells. Diethanolamine increases interleukin 6 (IL-6), IL-8, interferon γ-induced protein 10 kDa (IP-10), growth-regulated oncogene (GRO), fractalkine, matrix metalloproteinase 1 (MMP-1), MMP-9 and MMP-10 (p < 0.05 for all), factors involved in acute inflammation and recruitment of monocytes, neutrophils and lymphocytes. Ethyl glucoside and tartrate decrease multiple cytokines, including RANTES and MCP-1 (p < 0.05 for all), which serve as chemotactic factors. Vaginal cells exhibit heightened inflammatory tone compared with cervical cells and macrophages, with a greater number of differentially expressed analytes after xenobiotic exposure. CONCLUSIONS: Xenobiotic metabolites present in the cervicovaginal space during pregnancy modify immune responses, unveiling potential pathways through which environmental exposures may contribute to the pathogenesis of cervical remodelling preceding preterm birth. Future work identifying xenobiotic sources and routes of exposure offers the potential to modify environmental risks to improve pregnancy outcomes.
Assuntos
Etanolaminas , Nascimento Prematuro , Tartaratos , Recém-Nascido , Gravidez , Feminino , Humanos , Xenobióticos , Citocinas/metabolismo , Epitélio , ImunidadeRESUMO
Neutrophil binding to vascular P- and E-selectin is the rate-limiting step in the recruitment of immune cells to sites of inflammation. Many diseases, including sickle cell anemia, post-myocardial infarction reperfusion injury, and acute respiratory distress syndrome are characterized by dysregulated inflammation. We have recently reported sialyl Lewisx analogues as potent antagonists of P- and E-selectin and demonstrated their in vivo immunosuppressive activity. A key component of these molecules is a tartrate diester that serves as an acyclic tether to orient the fucoside and the galactoside moiety in the required gauche conformation for optimal binding. The next stage of our study involved attaching an extended carbon chain onto one of the esters. This chain could be utilized to tether other pharmacophores, lipids, and contrast agents in the context of enhancing pharmacological applications through the sialyl Lewisx / receptor-mediated mechanism. Herein, we report our preliminary studies to generate a small library of tartrate based sialyl Lewisx analogues bearing extended carbon chains. Anionic charged chemical entities are attached to take advantage of proximal charged amino acids in the carbohydrate recognition domain of the selectin receptors. Starting with a common azido intermediate, synthesized using copper-catalyzed Huisgen 1,3-dipolar cycloadditions, these molecules demonstrate E- and P-selectin binding properties.
Assuntos
Selectina E , Selectina-P , Humanos , Selectina-P/metabolismo , Selectina E/metabolismo , Tartaratos , Antígeno Sialil Lewis X , Oligossacarídeos/química , Sítios de Ligação , Carbono , Inflamação , Adesão CelularRESUMO
The extraction of cannabinoids from the inflorescence and leaves of Cannabis sativa L. is gaining interest from researchers, in addition to addressing the under-utilization of the by-products in the stems and roots of the trees. The present study investigated the recovery of pectin from the left-over parts of hemp tress using an eco-friendly method with the aid of organic acids. Different cannabis cultivars-Chalotte's Angels (CHA) and Hang-Krarog (HKR)-were used as plant materials. The stems of both cannabis cultivars contained more pectin than the roots, and tartaric acid-aided extraction provided higher yields than from citric acid. Extracting the acid solution affected some characteristics, thereby differentiating the functional properties of the derived pectin. Extraction using tartaric acid provided pectin with a higher galacturonic acid content, whereas pectin with a higher methylation degree could be prepared using citric acid. The pectin samples extracted from the stems of CHA (P-CHA) and HKR (P-HKR) had low methoxyl pectin. P-CHA had better free radical scavenging capability, whereas P-HKR showed more potent reducibility. Considering the functional properties, P-CHA showed greater emulsion formability and foaming activity, whereas P-HKR possessed a better thickening effect. The present work suggests the feasible utilization of P-CHA and P-HKR as food additives with bioactivity.
Assuntos
Cannabis , Pectinas , Extratos Vegetais , Pectinas/química , Pectinas/isolamento & purificação , Cannabis/química , Extratos Vegetais/química , Ácido Cítrico/química , Folhas de Planta/química , Caules de Planta/química , Tartaratos/química , Raízes de Plantas/química , Ácidos Hexurônicos/química , Ácidos Hexurônicos/análiseRESUMO
Low molecular weight organic acids (LMWOAs) are important soil components and play a key role in regulating the geochemical behavior of heavy metal(loid)s. Biochar (BC) is a commonly used amendment that could change LMWOAs in soil. Here, four LMWOAs of oxalic acid (OA), tartaric acid (TA), malic acid (MA), and citric acid (CA) were evaluated for their roles in changing Cd and SB desorption behavior in contaminated soil with (S1-BC) or without BC (S1) produced from Paulownia biowaste. The results showed that OA, TA, MA, and CA reduced soil pH with rising concentrations, and biochar partially offset the pH reduction by LMWOAs. The LMWOAs reduced Cd desorption from the soil at low concentrations but increased Cd desorption at high concentrations, and CA was the most powerful in this regard. The LMWOAs had a similar effect on Sb desorption, and CA was the most effective species of LMWOAs. Adding BC to the soil affects Cd and Sb dynamics by reducing the Cd desorption but increasing Sb desorption from the soil and increasing the distribution coefficient (Kd) values of Cd but lowering the Kd values of Sb. This study helped understand the effects of LMWOAs on the geochemical behavior of Cd and Sb in the presence of biochar, as well as the potential risks of biochar amendment in enhancing Sb desorption from contaminated soil.
Assuntos
Carvão Vegetal , Metais Pesados , Poluentes do Solo , Solo , Carvão Vegetal/química , Poluentes do Solo/química , Metais Pesados/química , Solo/química , Peso Molecular , Concentração de Íons de Hidrogênio , Cádmio/química , Tartaratos/química , Malatos/química , Ácido Cítrico/química , Recuperação e Remediação Ambiental/métodos , Ácido Oxálico/química , Adsorção , Oryza/químicaRESUMO
Vitamin C is extensively used in cosmetic formulation, howbeit stability is the supreme demerit that limits its use in beautifying products. Numerous techniques are being employed to inhibit the degradation of vitamin C caused by formulation components to facilitate the use in skin rejuvenating products. Diverse materials are being exercised in formulation to stabilize the ascorbic acid and ingredients selected in this formulation composition help for stabilization. The initial stable prototype is developed and further optimization is accomplished by applying the design of experiment tools. The stable pharmaceutical formulations were evaluated for the evaluation parameters and designated as two optimized formulations. The analytical method for the assay of ascorbic acid from the United States pharmacopeia and the related substance method from European pharmacopeia has been modified to be used for cream formulation. The DoE design exhibited that the stability of formulation is impacted by citric acid and tartaric acid but not by propylene glycol and glycerin. The analysis results of topical formulations for the evaluation parameter exhibited satisfactory results. The in-vitro release study method has been developed, optimized, and validated to fit the analysis. The in-vitro studies have been performed for selected compositions and both the formulation has similar kinds of release patterns. The stability study as per ICH guidelines exhibited that the product is stable for accelerated, intermediate, and room-temperature storage conditions. The optimized formulation shows constant release and permeation of ascorbic acid through the skin. The formulation with the combinations of citric acid, tartaric acid, and tocopherol is more stable and the degradation of vitamin C has been reduced significantly. The beaucoup strategies in the unique composition help to protect the degradation by inhibiting the multitudinous degradation pathways.
Assuntos
Ácido Ascórbico , Química Farmacêutica , Estabilidade de Medicamentos , Ácido Ascórbico/química , Química Farmacêutica/métodos , Tartaratos/química , Ácido Cítrico/química , Composição de Medicamentos/métodos , Excipientes/químicaRESUMO
BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important pathogen, characterized by its genetic and antigenic variation. The PRRSV vaccine is widely used, however, the unsatisfied heterologic protection and the risk of reverse virulence raise the requirement to find some new anti-PRRSV strategies for disease control. Tylvalosin tartrate is used to inhibit PRRSV in the field non-specifically, however, the mechanism is still less known. METHODS: The antiviral effects of Tylvalosin tartrates from three producers were evaluated in a cell inoculation model. Their safety and efficacy concentrations, and effecting stage during PRRSV infection were analyzed. And, the Tylvalosin tartrates regulated genes and pathways which are potentially related to the anti-viral effect were further explored by using transcriptomics analysis. Last, the transcription level of six anti-virus-related DEGs was selected to confirm by qPCR, and the expression level of HMOX1, a reported anti-PRRSV gene, was proved by western blot. RESULTS: The safety concentrations of Tylvalosin tartrates from three different producers were 40 µg/mL (Tyl A, Tyl B, and Tyl C) in MARC-145 cells and 20 µg/mL (Tyl A) or 40 µg/mL (Tyl B and Tyl C) in primary pulmonary alveolar macrophages (PAMs) respectively. Tylvalosin tartrate can inhibit PRRSV proliferation in a dose-dependent manner, causing more than 90% proliferation reduction at 40 µg/mL. But it shows no virucidal effect, and only achieves the antiviral effect via long-term action on the cells during the PRRSV proliferation. Furthermore, GO terms and KEGG pathway analysis was carried out based on the RNA sequencing and transcriptomic data. It was found that the Tylvalosin tartrates can regulate the signal transduction, proteolysis, and oxidation-reduction process, as well as some pathways such as protein digestion and absorption, PI3K-Akt signaling, FoxO signaling, and Ferroptosis pathways, which might relate to PRRSV proliferation or host innate immune response, but further studies still need to confirm it. Among them, six antivirus-related genes HMOX1, ATF3, FTH1, FTL, NR4A1, and CDKN1A were identified to be regulated by Tylvalosin tartrate, and the increased expression level of HMOX1 was further confirmed by western blot. CONCLUSIONS: Tylvalosin tartrate can inhibit PRRSV proliferation in vitro in a dose-dependent manner. The identified DEGs and pathways in transcriptomic data will provide valuable clues for further exploring the host cell restriction factors or anti-PRRSV target.
Assuntos
Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Suínos , Animais , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Vírus da Síndrome Respiratória e Reprodutiva Suína/metabolismo , Antivirais/farmacologia , Antivirais/metabolismo , Tartaratos/metabolismo , Tartaratos/farmacologia , Transcriptoma , Fosfatidilinositol 3-Quinases/metabolismo , Fosfatidilinositol 3-Quinases/farmacologia , Macrófagos Alveolares , Replicação ViralRESUMO
Olefin chemistry, through pericyclic reactions, polymerizations, oxidations, or reductions, has an essential role in the manipulation of organic matter. Despite its importance, olefin synthesis still relies largely on chemistry introduced more than three decades ago, with metathesis being the most recent addition. Here we describe a simple method of accessing olefins with any substitution pattern or geometry from one of the most ubiquitous and variegated building blocks of chemistry: alkyl carboxylic acids. The activating principles used in amide-bond synthesis can therefore be used, with nickel- or iron-based catalysis, to extract carbon dioxide from a carboxylic acid and economically replace it with an organozinc-derived olefin on a molar scale. We prepare more than 60 olefins across a range of substrate classes, and the ability to simplify retrosynthetic analysis is exemplified with the preparation of 16 different natural products across 10 different families.
Assuntos
Alcenos/química , Alcenos/síntese química , Produtos Biológicos/química , Produtos Biológicos/síntese química , Ácidos Carboxílicos/química , Alcenos/classificação , Amidas/química , Produtos Biológicos/classificação , Dióxido de Carbono/química , Dióxido de Carbono/isolamento & purificação , Catálise , Ferro/química , Níquel/química , Oxirredução , Policetídeos/síntese química , Policetídeos/química , Especificidade por Substrato , Tartaratos/síntese química , Tartaratos/química , Zinco/químicaRESUMO
A new eco-friendly catalytic system is devised for C-C bond formation through Suzuki coupling, using an impressive nanocatalyst (Fe3O4/L-(+)-tartaric acid/Pd-NPs). It contains immobilized palladium (0) onto magnetite nanoparticles, stabilized by tartaric acid, and is characterized by FT-IR, XRD, EDS, SEM, TEM, TGA, and VSM. The catalyst is used in an efficient synthesis of biaryls in EtOH/H2O (1:1), in the presence of K2CO3. Our Fe3O4/tartaric acid/Pd-NPs exhibit magnetic recoverability and reusability for five cycles without measurable loss of its activity.
Assuntos
Nanopartículas de Magnetita , Nanopartículas de Magnetita/química , Espectroscopia de Infravermelho com Transformada de Fourier , Tartaratos , CatáliseRESUMO
Most heavy metals and industrial chemicals such as nicotine and lead cause harm to the reproduction process through a decrease in sperm motility, fertilization process, and sperm binding to the oocyte. Salvia officinalis L. (sage) has been reported to enhance serum testosterone levels and other certain biochemical enzymes. Thus, the current study is aimed at evaluating the potential health benefits of S. officinalis L. methanol extract on lead and nicotine hydrogen tartrate-induced sperm quality degeneration in male rats and also identifying some of the non-polar volatile bioactive compounds that might be attributed to the bioactivity of S. officinalis extract using gas chromatography-mass spectrometry (GC/MS). In the study, fifty-four mature male albino rats of about 220-250â g [were divided randomly and equally into 9â groups (n=6)]. Sperm quality degeneration was induced through the oral administration of 1.5â g/L of lead acetate in drinking water or peritoneal injection of 0.50â mg/kg (animal weight) nicotine hydrogen tartrate for sixty days. Two doses (200 & 400â mg/kg b.w.) of S. officinalis L. were used. The rats were anesthetized after the experimental period and then sacrificed. Blood samples were collected while the epididymis, testicle, and accessory sex organs (prostates and seminal vesical) were taken for histopathological studies. Twelve major compounds were identified through the GC/MS analysis of S. officinalis L. methanol extract. Lead and nicotine toxicity had a great effect on the rats' sperm quality causing a significant (p<0.05) decrease in the quantity of sperm and sperm motility as well as an upsurge in the abnormalities of the sperm and a reduction in the length & diameter of seminiferous tubules and size & weight of sexual organs (accessory sex glands, epididymis, and testis). The administration of S. officinalis L. methanol extract, however, had a positive impact on the sexual organ weights, semen quality & quantity, and rats' fertility, thus, ameliorating the adversative effects of both lead and nicotine. Further evaluation and isolation of the bioactive components are recommended as potential drug leads.
Assuntos
Metanol , Salvia officinalis , Ratos , Masculino , Animais , Nicotina/farmacologia , Análise do Sêmen , Tartaratos/farmacologia , Ratos Wistar , Contagem de Espermatozoides/métodos , Motilidade dos Espermatozoides , Sementes , Espermatozoides , Extratos Vegetais/farmacologiaRESUMO
Here, we report a new version of the extended Rate Constants Distribution (RCD) model for metal ion sorption, which includes complex-formation equilibria. With the RCD-complex model, one can predict sorbent performance in the presence of complexing agents using data on metal ion sorption from ligand-free solutions and a set of coefficients for sorption rate constants of different ionic species. The RCD-complex model was applied to breakthrough curves of Cu(II) sorption from acetate and tartrate solutions on polyethyleneimine (PEI) monolith cryogel at different flow rates and ionic speciation. We have shown that, despite the lower stability of Cu(II)-acetate complex, at high flow rates, acetate has a more pronounced negative effect on sorption kinetics than tartrate. The RCD model was successfully used to predict the shape of the breakthrough curves at an arbitrary acetate concentration but failed to predict Cu(II) sorption from tartrate solutions in a broad range of ligand concentrations. Since a twofold increase in sorption capacity was observed at low tartrate concentrations, the latter fact was related to an alteration in the sorption mechanism of Cu(II)-ions, which depended on Cu(II) ionic speciation. The obtained results emphasize the importance of information about sorption kinetics of different ionic forms for the optimization of sorption filter performance in the presence of complexing agents.
Assuntos
Criogéis , Polietilenoimina , Cinética , Tartaratos , Concentração de Íons de Hidrogênio , Metais , Íons , Acetatos , Adsorção , Cobre , SoluçõesRESUMO
A novel Python-based open-source optimization framework, namely Pyomo (Python optimization modeling objects), alongside a conventional optimization method, RSM (response surface methodology), was utilized to determine the optimal operating conditions of an alternating current-powered electrocoagulation (ACPE) process for nickel removal. In this regard, four mutable operating factors, current density (5-9 mA/cm2), initial nickel concentration (200-400 mg/L), initial pH of the solution (5-9), and electrolysis time (30-60 min), along with a fixed amount of an additional eco-friendly substance, Tartaric Acid (155 mg/L) were considered. Metal removal efficiency (OF1) and operating costs (OF2) were monitored and evaluated as objective functions with the aim of maximization and minimization, respectively. Experiments were conducted according to the central composite design (CCD), and validation outcomes established a reasonable agreement between the predicted models and the experimental data. The multi-objective optimization process yielded two sets of 30-optimal-solution obtained through Pyomo and RSM. Accordingly, the proposed solutions by the Pyomo were found to be more flexible and eclectic, supplying the local decision maker(s) with a diverse spectrum of optimal operating conditions. Adding TA was also effective in reducing electrical energy consumption by up to 46%.
Assuntos
Níquel , Poluentes Químicos da Água , Eletrocoagulação , Eletrólise , TartaratosRESUMO
A smart temperature stimuli-driven multiplex photoelectrochemical (PEC) assay was constructed for antibiotic resistance genes (ARGs) detection, where the stimuli-responsive gatekeeping by regulating the alternative release of "cargo" allowed for the simultaneous detection of multiple tetracycline resistance gene, using tetA (TDNA1) and tetC (TDNA2) as the model. Dual temperature-responsive nanoassemblies were embedded in the PEC bioassay as signal DNA tages: one thermoresponsive polymer (poly(N-isopropylacrylamide), PNIPAM)-capped mesoporous silica nanoparticles (MSN) with loading the "cargo" of HgO nanoparticles as signal DNA1 tags (SDNA1-PNIPAM@MSN@HgONPs) and the other antimony tartrate (SbT)-anchored silica nanospheres as signal DNA2 tags (SDNA2-SbT@SiO2NSs). At 20 °C, below the lower critical solution temperature (LCST) of PNIPAM, the "gatekeeper" PNIPAM in SDNA1-PNIPAM@MSN@HgONPs was in an ON state, igniting Hg2+ release through the pore of SiO2. While at above LCST (40 °C), it was in an OFF state. Likewise, the thermo-dependent dissociation of SbT endowed the grafted SDNA2 tags switching from the OFF (at 20 °C) to ON state (at 40 °C), igniting SbO+ release. The released Hg2+ and SbO+ triggered the amplified photocurrents due to the structure evolution of the photoactive layer into HgS/ZnS or Sb2S3/ZnS heterostructure, thus achieving sensitive detection of multiple ARGs: tetA, tetC, tetG, tetM, tetO, tetZ, tetX, and tetW. Combined with heat map analysis, rapid screening of the ARGs profiles in 12 samples could be realized. This bioassay is simple and accessible for multiple genes analysis with the detection limit down to 0.50 nM. And it was successfully applied for measuring tetracycline ARGs in real sludge samples.
Assuntos
Mercúrio , Nanosferas , Antibacterianos/análise , Antibacterianos/farmacologia , Antimônio , Bioensaio , Polímeros , Esgotos , Dióxido de Silício/química , Sulfetos , Tartaratos , Temperatura , Tetraciclina , Compostos de ZincoRESUMO
The enolase superfamily (ENS) has served as a paradigm for understanding how enzymes that share a conserved structure, as well as a common partial reaction (i.e., metal-assisted, Brønsted base-catalyzed enol(ate) formation), evolved from a common progenitor to catalyze mechanistically diverse reactions. Enzymes of the mandelate racemase (MR)-subgroup of the ENS share interdigitating loops between adjacent, 2-fold symmetry-related protomers of the tightly associated homodimers that comprise their quaternary structures. For the MR-subgroup members MR and d-tartrate dehydratase (TarD), the tip of the loop contributes a binding determinant to the adjacent active site (i.e., Leu 93 and Lys 102, respectively). To assess the role of Leu 93 of MR in substrate specificity and catalysis, we constructed L93 variants bearing hydrophobic (L93A, L93F, and L93W), polar neutral (L93N), acidic (L93D), or basic (L93K and L93R) residues at position 93. Gel filtration-HPLC revealed that wild-type MR and all L93 MR variants, apart from L93R MR (dimeric), were tetrameric in solution. The catalytic efficiency (kcat/Km) was reduced in the RâS and SâR reaction directions for all variants, primarily due to reduced turnover (kcat). Substitution of Leu 93 by Lys or Arg to mimic Lys 102 of TarD enhanced the binding of malate and tartrate, with meso- and d-tartrate exhibiting linear mixed-type inhibition of L93K MR. Despite the striking 500-fold increase in the binding affinity of d-tartrate, relative to (S)-mandelate, L93K MR exhibited no TarD activity. MD simulations suggested that the failure of L93K MR to catalyze α-deprotonation (i.e., H-D exchange) arises from inappropriate positioning of the Brønsted base (Lys 166). Thus, a change in binding determinant on the interdigitating loop can play a significant role in governing substrate specificity within the ENS, but does not necessarily confer 'new' catalytic activity despite similarities in catalytic machinery.
Assuntos
Racemases e Epimerases , Tartaratos , Sítios de Ligação , Catálise , Hidroliases/química , Cinética , Modelos Moleculares , Racemases e Epimerases/genética , Racemases e Epimerases/metabolismo , Especificidade por SubstratoRESUMO
BACKGROUND AND AIMS: Soil phosphorus (P) deficiency and salinity are constraints to crop productivity in arid and semiarid regions. Salinity may weaken the effect of P fertilization on plant growth. We investigated the interactive effects of soil P availability and salinity on plant growth, P nutrition and salt tolerance of two alfalfa (Medicago sativa) cultivars. METHODS: A pot experiment was carried out to grow two cultivars of alfalfa in a loess soil under a combination of different rates of added P (0, 40, 80 and 160 mg P kg-1 soil as monopotassium phosphate) and sodium chloride (0, 0.4, 0.8 and 1.6 g NaCl kg-1 soil). Plant biomass, concentrations of P ([P]), sodium ([Na]) and potassium ([K]) were determined, and rhizosheath carboxylates were analysed. KEY RESULTS: There were significant interactions between soil P availability and salinity on some, but not all, of the parameters investigated, and interactions depended on cultivar. Plant growth and P uptake were enhanced by P fertilization, but inhibited by increased levels of salinity. Increasing the salinity resulted in decreased plant P-uptake efficiency and [K]/[Na]. Only soil P availability had a significant effect on the amount of tartrate in the rhizosheath of both cultivars. CONCLUSIONS: Increased salinity aggravated P deficiency. Appropriate application of P fertilizers improved the salt tolerance of alfalfa and increased its productivity in saline soils.
Assuntos
Medicago sativa , Salinidade , Fertilização , Fósforo , Raízes de Plantas , Sódio , Solo , Tartaratos/farmacologiaRESUMO
A sulphate-reducing magnetotactic bacterium, designated strain FSS-1T, was isolated from sediments and freshwater of Suwa Pond located in Hidaka, Saitama, Japan. Strain FSS-1T was a motile, Gram-negative and curved rod-shaped bacterium that synthesizes bullet-shaped magnetite (Fe3O4) nanoparticles in each cell. Strain FSS-1T was able to grow in the range of pH 6.5-8.0 (optimum, pH 7.0), 22-34 °C (optimum, 28 °C) and with 0-8.0 g l-1 NaCl (optimum, 0-2.0 g l-1 NaCl). Strain FSS-1T grew well in the presence of 50 µM ferric quinate as an iron source. The major fatty acids were anteiso-C15â:â0, iso-C15â:â0 and anteiso-C17â:â0. The major menaquinone was MK-7 (H2). Strain FSS-1T contained desulfoviridin, cytochrome c 3 and catalase, but did not contain oxidase. Strain FSS-1T used fumarate, lactate, pyruvate, malate, formate/acetate, succinate, tartrate, ethanol, 1-propanol, peptone, soytone and yeast extract as electron donors, while the strain used sulphate, thiosulphate and fumarate as electron acceptors. Fumarate was fermented in the absence of electron acceptors. Analysis of the 16S rRNA gene sequence showed that strain FSS-1T is a member of the genus Fundidesulfovibrio. The gene sequence showed 96.7, 95.0, 92.0, 91.2 and 91.4% similarities to the most closely related members of the genera Fundidesulfovibrio putealis B7-43T, Fundidesulfovibrio butyratiphilus BSYT, Desulfolutivibrio sulfoxidireducens DSM 107105T, Desulfolutivibrio sulfodismutans ThAc01T and Solidesulfovibrio magneticus RS-1T, respectively. The DNA G+C content of strain FSS-1T was 67.5 mol%. The average nucleotide identity value between strain FSS-1T and F. putealis B7-43T was 80.7â%. Therefore, strain FSS-1T represents a novel species within the genus Fundidesulfovibrio, for which the name Fundidesulfovibrio magnetotacticus sp. nov. is proposed (=JCM 32405T=DSM 110007T).
Assuntos
Sulfatos , Tartaratos , 1-Propanol , Técnicas de Tipagem Bacteriana , Composição de Bases , Catalase/genética , Citocromos c/genética , DNA Bacteriano/genética , Etanol , Ácidos Graxos/química , Óxido Ferroso-Férrico , Formiatos , Fumaratos , Sulfito de Hidrogênio Redutase/genética , Ferro , Lactatos , Malatos , Nucleotídeos , Peptonas , Filogenia , Lagoas , Piruvatos , Ácido Quínico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Cloreto de Sódio , Succinatos , Tiossulfatos , Vitamina K 2RESUMO
Oxygen vacancies play a vital role in the catalytic activity of layered double hydroxide (LDH) catalysts in wastewater treatment. However, the mechanism of oxygen vacancy-mediated LDH-activated oxygen to produce reactive oxygen species (ROS) still lacks a reasonable explanation. In this work, a tartrate-modified CuCoFe-LDH (CuCoFe/Tar-LDH) with abundant oxygen vacancies was designed, which can efficiently degrade nitrobenzene (NB) under room conditions. The technical energy consumption is 0.011 kW h L-1. According to the characterization and calculation results, it is proposed that oxygen vacancies are formed because of the oxygen deficiency which is caused by the reduction of the energy between the metal ion and oxygen, and the metal ion transitions to a lower state. Compared with CuCoFe-LDH, the oxygen vacancy formation energy of CuCoFe/Tar-LDH decreased from 1.98 to 1.13 eV. The O2 bond length adsorbed on the oxygen vacancy is 1.27 Å, close to the theoretical length of superoxide radicals (â¢O2-) (1.26 Å). Radical trapping experiments and electron spin-resonance spectroscopy spectrum prove that â¢O2- is an important precursor of â¢OH. This work is dedicated to the in-depth exploration of the oxygen vacancy-mediated CuCoFe/Tar-LDH catalyst activation mechanism for molecular oxygen and the conversion relationship between ROS.
Assuntos
Oxigênio , Superóxidos , Hidróxidos/química , Nitrobenzenos , Oxigênio/química , Espécies Reativas de Oxigênio , TartaratosRESUMO
Tartaric acid and mucic acid are dicarboxylic acids (DCAs), a substance class often found in atmospheric aerosols and cloud droplets. The hydroxyl radical (â¢OH)-induced oxidation in the aqueous phase is known to be an important loss process of organic compounds such as DCAs. However, the study of â¢OH kinetics of DCAs in the aqueous phase is still incomplete. In the present study, the rate constants of the â¢OH reactions of tartaric acid and mucic acid in the aqueous phase were determined by the thiocyanate competition kinetics method as a function of temperature and pH. The following T-dependent Arrhenius expressions (in units of L mol-1 s-1) were first derived for the â¢OH reactions with tartaric acidâk(T, H2A) = (3.3 ± 0.1) × 1010 exp[(-1350 ± 110 K)/T], k(T, HA-) = (3.6 ± 0.1) × 1010 exp[(-580 ± 110 K)/T], and k(T, A2-) = (3.3 ± 0.1) × 1010 exp[(-1190 ± 170 K)/T]âas well as mucic acidâk(T, H2A) = (2.2 ± 0.1) × 1010 exp[(-1140 ± 150 K)/T], k(T, HA-) = (4.8 ± 0.1) × 1010 exp[(-1280 ± 170 K)/T], and k(T, A2-) = (2.1 ± 0.1) × 1010 exp[(-970 ± 70 K)/T]. A general trend of the â¢OH rate constant is found as kA2- > kHA- > kH2A. The pH- and temperature-dependent rate constants of the OH radical reactions allow an accurate description of the source and sink processes in the tropospheric aqueous phase.