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1.
Environ Sci Process Impacts ; 26(3): 519-529, 2024 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-38344926

RESUMO

The environmental and climatic factors dictating atmospheric mercury (Hg) uptake by foliage and accumulation within the forest floor are evaluated across six mountain sites, South Korea, using Hg concentration and Hg stable isotope analyses. The isotope ratios of total gaseous Hg (TGM) at six mountains are explained by local anthropogenic Hg emission influence and partly by mountain elevation and wind speed. The extent to which TGM is taken up by foliage is not dependent on the site-specific TGM concentration, but by the local wind speed, which facilitates TGM passage through dense deciduous canopies in the Korean forests. This is depicted by the significant positive relationship between wind speed and foliage Hg concentration (r2 = 0.92, p < 0.05) and the magnitude of δ202Hg shift from TGM to foliage (r2 = 0.37, p > 0.05), associated with TGM uptake and oxidation by foliar tissues. The litter and topsoil Hg concentrations and isotope ratios reveal relationships with a wide range of factors, revealing lower Hg level and greater isotopic fractionation at sites with low elevation, high wind speed, and high mean warmest temperature. We attribute this phenomenon to active TGM re-emission from the forest floor at sites with high wind speed and high temperature, caused by turnover of labile organic matter and decomposition. In contrast to prior studies, we observe no significant effect of precipitation on forest Hg accumulation but precipitation appears to reduce foliage-level Hg uptake by scavenging atmospheric Hg species available for stomata uptake. The results of this study would enable better prediction of future atmospheric and forest Hg influence under climate change.


Assuntos
Mercúrio , Mercúrio/análise , Monitoramento Ambiental , Florestas , Isótopos/análise , Gases/análise
2.
Environ Sci Technol ; 58(5): 2564-2573, 2024 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-38278139

RESUMO

The role of pH in sequestration of Cr(VI) by zerovalent magnesium (ZVMg) was characterized by global fitting of a kinetic model to time-series data from unbuffered batch experiments with varying initial pH values. At initial pH values ranging from 2.0 to 6.8, ZVMg (0.5 g/L) completely reduced Cr(VI) (18.1 µM) within 24 h, during which time pH rapidly increased to a plateau value of ∼10. Time-series correlation analysis of the pH and aqueous Cr(VI), Cr(III), and Mg(II) concentration data suggested that these conditions are controlled by combinations of reactions (involving Mg0 oxidative dissolution and Cr(VI) sequestration) that evolve over the time course of each experiment. Since this is also likely to occur during any engineering applications of ZVMg for remediation, we developed a kinetic model for dynamic pH changes coupled with ZVMg corrosion processes. Using this model, the synchronous changes in Cr(VI) and Mg(II) concentrations were fully predicted based on the Langmuir-Hinshelwood kinetics and transition-state theory, respectively. The reactivity of ZVMg was different in two pH regimes that were pH-dependent at pH < 4 and pH-independent at the higher pH. This contrasting pH effect could be ascribed to the shift of the primary oxidant of ZVMg from H+ to H2O at the lower and higher pH regimes, respectively.


Assuntos
Cromatos , Poluentes Químicos da Água , Cromatos/química , Magnésio , Ferro/química , Poluentes Químicos da Água/análise , Cromo/análise , Cromo/química , Cinética , Concentração de Íons de Hidrogênio , Adsorção
3.
J Pers Med ; 13(8)2023 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-37623447

RESUMO

Positive airway pressure (PAP) is an important treatment tool for patients with moderate and severe obstructive sleep apnea (OSA), and adherence to PAP significantly affects treatment outcomes. Disease severity, adverse effects, and psychosocial factors are known to predict medication adherence. Cephalometric parameters have been reported to positively correlate with upper airway collapse. However, research on the correlation between these cephalometric parameters and PAP adherence remains insufficient. This study aimed to identify this relationship. This study included 185 patients with OSA who were prescribed PAP. Polysomnography (PSG) was performed to diagnose OSA, and paranasal sinus computed tomography (PNS CT) was performed to check for comorbidities of the upper airway. In addition, cephalometric parameters such as the hyoid-posterior nasal spine (H-PNS), posterior nasal spine-mandibular plane (PNS-MP), and hyoid-mandibular plane (H-MP) were measured in the midsagittal and axial CT views. Adherence was evaluated 3-12 months after the PAP prescription. A total of 136 patients were PAP-adherent, and 49 were nonadherent. There were more males in the adherent group and a higher average height in the adherent group. The PSG results showed that the apnea-hypopnea index (AHI), respiratory disturbance index (RDI), oxygen desaturation index (ODI), arousal index (AI), rapid eye movement (REM) AHI, and supine AHI were significantly higher, and the lowest oxygen saturation was lower in the adherent group. In the analysis of covariance (ANCOVA) model adjusted for sex and height, among the cephalometric parameters, H-MP was significantly longer in the adherent group (p = 0.027), and H-PNS showed a longer tendency (p = 0.074). In the logistic regression analysis model, the odds ratio (OR) and 95% confidence intervals (95% CI) of adherence and severe OSA in the third tertile compared to the first tertile of H-MP were 2.93 (1.25-6.86) and 4.00 (1.87-8.56). In the case of H-PNS, they were 2.58 (1.14-5.81) and 4.86 (2.24-10.54), respectively. This study concluded that an inferiorly placed hyoid bone in adult patients is associated with PAP adherence and disease severity.

4.
Environ Sci Technol ; 56(15): 10808-10817, 2022 08 02.
Artigo em Inglês | MEDLINE | ID: mdl-35852377

RESUMO

We coupled compound-specific isotopic analyses of nitrogen (N) in amino acids (δ15NGlu, δ15NPhe) and mercury stable isotopes (δ202Hg, Δ199Hg) to quantify ecological traits governing the concentration, variability, and source of Hg in largemouth bass (LB) and pike gudgeon (PG) across four rivers, South Korea. PG displayed uniform Hg concentration (56-137 ng/g), trophic position (TPcorrected; 2.6-3.0, n = 9), and N isotopes in the source amino acid (δ15NPhe; 7-13‰), consistent with their specialist feeding on benthic insects. LB showed wide ranges in Hg concentration (45-693 ng/g), TPcorrected (2.8-3.8, n = 14), and δ15NPhe (1.3-16‰), reflecting their opportunistic feeding behavior. Hg sources assessed using Hg isotopes reveal low and uniform Δ199Hg in PG (0.20-0.49‰), similar to Δ199Hg reported in sediments. LB displayed site-specific δ202Hg (-0.61 to -0.04‰) and Δ199Hg (0.53-1.09‰). At the Yeongsan River, LB displayed elevated Δ199Hg and low δ15NPhe, consistent with Hg and N sourced from the atmosphere. LB at the Geum River displayed low Δ199Hg and high δ15NPhe, both similar to the isotope values of anthropogenic sources. Our results suggest that a specialist fish (PG) with consistent ecological traits and Hg concentration is an effective bioindicator species for Hg. When accounting for Hg sources, however, LB better captures site-specific Hg sources.


Assuntos
Mercúrio , Poluentes Químicos da Água , Animais , Monitoramento Biológico , Monitoramento Ambiental , Peixes/metabolismo , Isótopos , Mercúrio/análise , Isótopos de Mercúrio/análise , Nitrogênio/análise , Isótopos de Nitrogênio/análise , Poluentes Químicos da Água/análise
5.
Sci Total Environ ; 814: 152598, 2022 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-34958842

RESUMO

Nearshore systems play an important role as mercury (Hg) sources to the open ocean and to human health via fish consumption. The nearshore system along East Asia is of particular concern given the rapid industrialization, which contributes to significant anthropogenic Hg emissions and releases. We used Hg stable isotopes to characterize Hg sources in the sediment and fish along the west coast of Korea, located at the northeast of the East China Sea. The Hg isotope ratios of the west coast sediments (δ202Hg; -0.89 to -0.27‰, Δ199Hg; -0.04 to 0.14‰) were statistically similar with other nearshore sediments (δ202Hg; -0.99 to -0.30‰, Δ199Hg; -0.04 to 0.19‰) and overlapped with the industrial Hg source end-member (δ202Hg; -0.5‰, Δ199Hg; 0.01‰) estimated from the Chinese marginal seas. Using a ternary mixing model, we estimated that industrial Hg sources contribute 83-97% in the west coast of Korea, and riverine and atmospheric Hg sources play minor roles in the Korean west coast and the Chinese marginal seas. The comparison between Hg isotope ratios of the sediment and nearshore fish revealed that the fish in the most west coast sites are exposed to MeHg produced in the sediment. At a few southwest coast sites, external MeHg produced in rivers and the open ocean water column appears to be more important as a source in fish. This is supported by much higher δ202Hg (0.74‰; similar to oceanic fish) and lower δ202Hg (-0.71‰; similar to riverine sources) compared to fish collected from other west coast sites influenced by sedimentary MeHg. The substantial Hg contributions from industrial activities suggest the national policies regulating anthropogenic Hg releases can directly mitigate human Hg exposure originating via local fish consumption. This study contributes to the growing regional and global inventories of Hg fluxes and sources exported into coastal oceans.


Assuntos
Mercúrio , Poluentes Químicos da Água , Animais , Monitoramento Ambiental , Sedimentos Geológicos , Humanos , Isótopos , Mercúrio/análise , Isótopos de Mercúrio/análise , Oceanos e Mares , Poluentes Químicos da Água/análise
6.
Environ Monit Assess ; 192(1): 41, 2019 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-31834560

RESUMO

To evaluate the pretreatment processing for Cd isotope analysis of environmental samples, acid extractions and total digestions were examined with various environmental certified reference materials. Four certified reference material samples, including fly ash, polluted soil, domestic sludge, and industrial sludge, were digested by six different acid extraction and total digestion methods, and then Cd was separated to remove other matrix by anion-exchange column. The Cd recovery rates of the acid extraction methods were 2.6-89.1%, while those of the total digestion methods were 21.6-88.7%. In these results, the Cd recovery rates were dependent on the sample type. More than 80% of the Cd in the polluted soil and domestic sludge samples could be recovered regardless of the decomposition method, except one method. On the other hand, the Cd recovery rate from fly ash was low when total digestion was performed using a HF mixture, and the recovery rate by total digestion methods for industrial sludge was higher than that by acid extraction. In our results, Cd isotope ratios tended to be more positive by increasing the Cd recovery rates, suggesting that the light isotope of Cd was decomposed preferentially during the decomposition procedures. However, when more than 80% of the Cd in the samples was recovered, the Cd isotope ratios were determined to be similar. This indicated that at least 80% of the Cd should be recovered from environmental samples to accurately measure the Cd isotopic ratio of environmental samples.


Assuntos
Cádmio/análise , Monitoramento Ambiental , Poluentes Ambientais/análise , Ácidos , Cinza de Carvão , Isótopos , Esgotos
7.
Environ Geochem Health ; 41(6): 2443-2458, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31016607

RESUMO

Among the results of community health impact assessments completed in 2014, residents of the Indae abandoned metal mine area showed high average urinary concentrations of harmful arsenic (As), at 148.9 µg/L. The concentration of harmful As was derived as the sum of As(V), As(III), MMA, and DMA concentrations known to be toxic. In this area, mining hazard prevention work was not carried out and the pollution source was neglected, and the health effect of the residents due to arsenic exposure was concerned. We re-assessed As exposure levels and tried to identify exposure factors for residents of this area. Analysis of the soil, sediment, and river water to assess the association between the soil of the Indae abandoned metal mine area and the soil in residential areas confirmed a correlation between Pb and As concentrations in the soil. Since Pb and As behave similarly, the use of the stable Pb isotope ratio for assessment of the pollution source tracking was validated. In the 3-isotope plot (207/206Pb vs. 208/206Pb) of soil samples in this area, a stable Pb isotope ratio was located on the same trend line, which confirmed that the soil in the residential area was within the area of influence of the Indae abandoned metal mine. Therefore, we judged that the pollution source of As was the Indae abandoned metal mine. The results by As species were As (III) 1.45 µg/L, As (V) 0.74 µg/L, monomethylarsonic acid (MMA) 2.43 µg/L, dimethylarsinic acid (DMA) 27.63 µg/L, and arsenobetaine 88.62 µg/L. The urinary harmful As was 31.92 µg/L, much lower than the 148.9 µg/L reported in a 2014 survey, due to the implementation of a multi-regional water supply in November 2014 that restricted As exposure through drinking river water. However, concerns remain over chronic exposure to As because As in river water used for farming and in agricultural soil still exceeds environmental standards; thus, ongoing work to address hazards from former mining areas and continued environmental monitoring is necessary.


Assuntos
Arsênio/análise , Exposição Ambiental/análise , Mineração , Poluentes do Solo/análise , Poluentes Químicos da Água/análise , Adulto , Idoso , Agricultura , Arsênio/urina , Arsenicais/análise , Exposição Ambiental/efeitos adversos , Monitoramento Ambiental , Feminino , Água Doce/análise , Sedimentos Geológicos/análise , Humanos , Isótopos/análise , Chumbo/análise , Masculino , Pessoa de Meia-Idade , República da Coreia , Rios/química , Fatores Socioeconômicos , Inquéritos e Questionários , Abastecimento de Água
8.
Appl Microbiol Biotechnol ; 102(20): 8855-8866, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30128580

RESUMO

Agar is a major polysaccharide of red algal cells and is mainly decomposed into neoagarobiose by the co-operative effort of ß-agarases. Neoagarobiose is hydrolyzed into monomers, D-galactose and 3,6-anhydro-L-galactose, via a microbial oxidative process. Therefore, the enzyme, 1,3-α-3,6-anhydro-L-galactosidase (α-neoagarobiose/neoagarooligosaccharide hydrolase) involved in the final step of the agarolytic pathway is crucial for bioindustrial application of agar. A novel cold-adapted α-neoagarooligosaccharide hydrolase, Ahg786, was identified and characterized from an agarolytic marine bacterium Gayadomonas joobiniege G7. Ahg786 comprises 400 amino acid residues (45.3 kDa), including a 25 amino acid signal peptide. Although it was annotated as a hypothetical protein from the genomic sequencing analysis, NCBI BLAST search showed 57, 58, and 59% identities with the characterized α-neoagarooligosaccharide hydrolases from Saccharophagus degradans 2-40, Zobellia galactanivorans, and Bacteroides plebeius, respectively. The signal peptide-deleted recombinant Ahg786 expressed and purified from Escherichia coli showed dimeric forms and hydrolyzed neoagarobiose, neoagarotetraose, and neoagarohexaose into 3,6-anhydro-L-galactose and other compounds by cleaving α-1,3-glycosidic bonds from the non-reducing ends of neoagarooligosaccharides, as confirmed by thin-layer chromatography and mass spectrometry. The optimum pH and temperature for Ahg786 activity were 7.0 and 15 °C, respectively, indicative of its unique cold-adapted features. The enzymatic activity severely inhibited with 0.5 mM ethylenediaminetetraacetic acid was completely restored or remarkably enhanced by Mn2+ in a concentration-dependent manner, suggestive of the dependence of the enzyme on Mn2+ ions. Km and Vmax values for neoagarobiose were 4.5 mM and 1.33 U/mg, respectively.


Assuntos
Alteromonadaceae/enzimologia , Proteínas de Bactérias/química , Galactosidases/química , Alteromonadaceae/química , Alteromonadaceae/genética , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Estabilidade Enzimática , Galactosidases/genética , Galactosidases/metabolismo , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , Sinais Direcionadores de Proteínas , Alinhamento de Sequência , Temperatura
9.
Regul Toxicol Pharmacol ; 90: 9-21, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28782575

RESUMO

Agar, a heterogeneous polymer of galactose, is the main component of the cell wall of marine red algae. It is well established as a safe, non-digestible carbohydrate in Oriental countries. Although neoagarooligosaccharides (NAOs) prepared by the hydrolysis of agar by ß-agarase have been reported to exert various biological activities, the safety of these compounds has not been reported to date. For safety evaluation, NAOs containing mainly neoagarotetraose and neoagarohexaose were prepared from agar by enzymatic hydrolysis using ß-agarase DagA from Streptomyces coelicolor. Genotoxicity tests such as the bacterial reverse mutation assay, eukaryotic chromosome aberration assay, and in vivo micronucleus assay all indicated that NAOs did not exert any mutational effects. The toxicity of NAOs in rat and beagle dog models was investigated by acute, 14-day, and 91-day repeated oral dose toxicity tests. The results showed that NAO intake of up to 5,000 mg/kg body weight resulted in no significant changes in body weight, food intake, water consumption, hematologic and blood biochemistry parameters, organ weight, or clinical symptoms. Collectively, a no-observed-adverse-effect level of 5,000 mg/kg body weight/day for both male and female rats was established for NAO. These findings support the safety of NAO for possible use in food supplements and pharmaceutical and cosmetic products.


Assuntos
Ágar/toxicidade , Galactosídeos/toxicidade , Oligossacarídeos/toxicidade , Ágar/química , Animais , Peso Corporal , Linhagem Celular , Cosméticos/química , Cosméticos/toxicidade , Cricetulus , Suplementos Nutricionais/toxicidade , Cães , Feminino , Glicosídeo Hidrolases/química , Hidrólise , Masculino , Camundongos , Camundongos Endogâmicos ICR , Modelos Animais , Testes de Mutagenicidade/métodos , Nível de Efeito Adverso não Observado , Ratos , Ratos Sprague-Dawley
10.
Mar Drugs ; 15(4)2017 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-28333098

RESUMO

Neoagarooligosaccharides (NAOs), mainly comprising neoagarotetraose and neoagarohexaose, were prepared by hydrolyzing agar with ß-agarase DagA from Streptomyces coelicolor, and the anti-obesity and anti-diabetic effects of NAOs on high-fat diet (HFD)-induced obesity in mice were investigated after NAOs-supplementation for 64 days. Compared to the HFD group, the HFD-0.5 group that was fed with HFD + NAOs (0.5%, w/w) showed remarkable reduction of 36% for body weight gain and 37% for food efficiency ratios without abnormal clinical signs. Furthermore, fat accumulation in the liver and development of macrovesicular steatosis induced by HFD in the HFD-0.5 group were recovered nearly to the levels found in the normal diet (ND) group. NAOs intake could also effectively reduce the size (area) of adipocytes and tissue weight gain in the perirenal and epididymal adipose tissues. The increased concentrations of total cholesterol, triglyceride, and free fatty acid in serum of the HFD group were also markedly ameliorated to the levels found in serum of the ND group after NAOs-intake in a dose dependent manner. In addition, insulin resistance and glucose intolerance induced by HFD were distinctly improved, and adiponectin concentration in the blood was notably increased. All these results strongly suggest that intake of NAOs can effectively suppress obesity and obesity-related metabolic syndromes, such as hyperlipidemia, steatosis, insulin resistance, and glucose intolerance, by inducing production of adiponectin in the HFD-induced obese mice.


Assuntos
Fármacos Antiobesidade/farmacologia , Diabetes Mellitus/tratamento farmacológico , Hipoglicemiantes/farmacologia , Obesidade/tratamento farmacológico , Oligossacarídeos/farmacologia , Adipócitos/efeitos dos fármacos , Tecido Adiposo/efeitos dos fármacos , Animais , Glicemia/efeitos dos fármacos , Colesterol/metabolismo , Dieta Hiperlipídica , Intolerância à Glucose/tratamento farmacológico , Resistência à Insulina/fisiologia , Fígado/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fitoterapia/métodos , Extratos Vegetais/farmacologia , Triglicerídeos/metabolismo , Aumento de Peso/efeitos dos fármacos
11.
J Microbiol ; 53(7): 448-53, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26115993

RESUMO

A xylan-degrading bacterium, designated as MS5(T) strain, was isolated from soil collected from the Jeju Island, Republic of Korea. Strain MS5(T) was Gram-stain-positive, aerobic, and motile by polar flagellum. The major fatty acids identified in this bacterium were iso-C15:0 (32.3%), C16:0 (27.3%), and anteiso-C15:0 (10.2%). A similarity search based on the 16S rRNA gene sequence revealed that the strain belongs to the class Bacilli and shared the highest similarity with the type strains Bacillus beringensis BR035(T) (98.7%) and Bacillus korlensis ZLC-26(T) (98.6%) which form a coherent cluster in a neighbor-joining phylogenetic tree. The DNA G+C content of strain MS5(T) was 43.0 mol%. The major menaquinone was MK-7 and the diagnostic diamino acid in the cell-wall peptidoglycan was meso-diaminopimelic acid. The DNADNA relatedness values between strain MS5(T) and two closely related species, B. beringensis BR035(T) and B. korlensis ZLC-26(T), were less than 70%. DNA-DNA relatedness analysis and 16S rRNA sequence similarity, as well as phenotypic and chemotaxonomic characteristics suggest that the strain MS5(T) constitutes a novel Bacillus species, for which the name Bacillus coreaensis sp. nov. is proposed. The type strain is MS5(T) (=DSM25506(T) =KCTC13895(T)).


Assuntos
Bacillus/isolamento & purificação , Bacillus/metabolismo , Microbiologia do Solo , Xilanos/metabolismo , Bacillus/classificação , Bacillus/fisiologia , Composição de Bases , Ácido Diaminopimélico/análise , Ácidos Graxos/análise , Hidrólise , Fenótipo , Filogenia , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/análise
12.
FEMS Microbiol Lett ; 362(7)2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25761755

RESUMO

A putative agarase gene (agaH92) encoding a primary translation product (50.1 kDa) of 445 amino acids with a 19-amino-acid signal peptide and glycoside hydrolase 16 and RICIN superfamily domains was identified in an agarolytic marine bacterium, Pseudoalteromonas sp. H9 ( = KCTC23887). The heterologously expressed protein rAgaH92 in Escherichia coli had an apparent molecular weight of 51 kDa on SDS-PAGE, consistent with the calculated molecular weight. Agarase activity of rAgaH92 was confirmed by a zymogram assay. rAgaH92 hydrolyzed p-nitrophenyl-ß-D-galactopyranoside, but not p-nitrophenyl-α-D-galactopyranoside. The optimum pH and temperature for rAgaH92 were 6.0 and 45°C, respectively. It was thermostable and retained more than 85% of its initial activity after heat treatment at 50°C for 1 h. rAgaH92 required Fe(2+) for agarase activity and inhibition by EDTA was compensated by Fe(2+). TLC analysis, mass spectrometry and NMR spectrometry of the GST-AgaH71 hydrolysis products revealed that rAgaH92 is an endo-type ß-agarase, hydrolyzing agarose into neoagarotetraose and neoagarohexaose.


Assuntos
Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/metabolismo , Pseudoalteromonas/enzimologia , Pseudoalteromonas/genética , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Proteínas de Bactérias/metabolismo , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Escherichia coli/genética , Galactosídeos/metabolismo , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/isolamento & purificação , Ferro/metabolismo , Peso Molecular , Nitrofenilgalactosídeos/metabolismo , Oligossacarídeos/metabolismo , Sinais Direcionadores de Proteínas , Sefarose/metabolismo , Especificidade por Substrato
13.
Appl Biochem Biotechnol ; 175(2): 733-47, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25342256

RESUMO

An agarase gene (agaH71) was identified from Pseudoalteromonas hodoensis, an agar utilizing marine bacterium. The nucleotide sequence revealed that AgaH71 had significant homology to glycosyl hydrolase (GH) 16 agarases. agaH71 encodes a primary translation product (32.7 kDa) of 290 amino acids, including a 21-amino-acid signal peptide. The entire AgaH71 was expressed in a fused protein with glutathione-S-transferase (GST) at its N-terminal (GST-AgaH71) in Escherichia coli. Purified GST-AgaH71 had an apparent molecular weight of 59 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), which was consistent with the calculated molecular weight (58.7 kDa). Agarase activity of the purified protein was confirmed by zymogram assay. GST-AgaH71 could hydrolyze p-nitrophenyl-ß-D-galactopyranoside, but not p-nitrophenyl-α-D-galactopyranoside. The optimum pH and temperature for GST-AgaH71 were 6.0 and 45 °C, respectively. GST-AgaH71 retained more than 95 and 90 % of its initial activity at 40 and 45 °C after heat treatment for 60 min, respectively. The K m and V max for agarose were 28.33 mg/ml and 88.25 U/mg, respectively. GST-AgaH71 did not require metal ions for its activity, but severe inhibition by divalent metal ions was observed. Thin-layer chromatography (TLC) analysis, mass spectrometry, and nuclear magnetic resonance (NMR) spectrometry of the GST-AgaH71 hydrolysis products revealed that GST-AgaH71 is an endo-type ß-agarase that hydrolyzes agarose into predominantly neoagarotetraose and small proportions of neoagarobiose and neoagarohexaose.


Assuntos
Proteínas de Bactérias/química , Glicosídeo Hidrolases/química , Pseudoalteromonas/química , Sefarose/química , Proteínas de Bactérias/genética , Clonagem Molecular , Dissacarídeos/química , Escherichia coli/genética , Escherichia coli/metabolismo , Galactosídeos/química , Expressão Gênica , Glutationa Transferase/química , Glutationa Transferase/genética , Glicosídeo Hidrolases/genética , Hidrólise , Cinética , Peso Molecular , Oligossacarídeos/química , Fases de Leitura Aberta , Sinais Direcionadores de Proteínas , Pseudoalteromonas/enzimologia , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Especificidade por Substrato
14.
Appl Biochem Biotechnol ; 173(7): 1703-16, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24879592

RESUMO

A Gram-negative, aerobic, motile, rod-shaped, agarolytic bacterium, designated as H7, was isolated from a coastal seawater sample. This strain grows at pH 6.0-8.0, temperature of 15-40 °C, and at an NaCl concentration of 1-7% (w/v). Ubiquinone-8 was the predominant respiratory quinone, and the DNA G+C content was 45.82 mol%. Analysis of the 16S rRNA sequence suggests that strain H7 belongs to the genus Pseudoalteromonas. DNA-DNA hybridization analysis showed DNA relatedness of as low as 55.42 and 40.27% with its nearest phylogenetic neighbors Pseudoalteromonas atlantica IAM12927T and Pseudoalteromonas espejiana NCIMB2127T, respectively, which led us to name H7 Pseudoalteromonas hodoensis sp. nov. The type strain is H7T (=DSM25967T=KCTC23887T). An agarase (AgaA7) was purified to homogeneity from the cell-free culture broth of H7 through many steps of chromatography. Purified AgaA7 had an apparent molecular weight of 35 kDa, with a distinct NH2-terminal sequence of Ala-Asp-Ala-Thr-X-Pro (X, any amino acid) from the reported proteins, implying that it is a novel enzyme. The optimum pH and temperature for agarase activity were 7.0 and 45 °C, respectively. Thin-layer chromatography analysis, mass spectrometry, and enzyme assay using p-nitrophenyl-α/ß-D-galactopyranoside revealed that AgaA7 is both an exo- and endo-type ß-agarase that degrades agarose into neoagarotetraose, neoagarohexaose, and neoagarooctaose (minor).


Assuntos
Espaço Extracelular/enzimologia , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/metabolismo , Pseudoalteromonas/enzimologia , Pseudoalteromonas/isolamento & purificação , Água do Mar/microbiologia , Temperatura , Ágar/metabolismo , Estabilidade Enzimática , Glicosídeo Hidrolases/biossíntese , Hidrólise , Fenótipo , Filogenia , Pseudoalteromonas/classificação , Pseudoalteromonas/citologia , República da Coreia
15.
Environ Monit Assess ; 186(2): 781-9, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24197555

RESUMO

This study focused on Pb isotope ratios of sediments in areas around an abandoned mine to determine if the ratios can be used as a source tracer. For pretreatment, sediment samples were dissolved with mixed acids, and a multi-collector inductively coupled plasma mass spectrometer (MC-ICP-MS, Nu plasma II) was used to investigate the Pb isotopic composition of the samples. The measured isotope ratios were then corrected for instrumental mass fractionation by measuring the (203)Tl/(205)Tl ratio. Repeated measurements with the NIST SRM 981 reference material showed that the precision of all ratios was below 104 ppm (±2σ) for 50 ng/g. The isotope ratios ((207)Pb/(206)Pb) found were 0.85073 ± 0.0004~0.85373 ± 0.0003 for the main stream, while they were 0.83736 ± 0.0010 for the tributary and 0.84393 ± 0.0002 for the confluence. A binary mixing equation for isotope ratios showed that the contributions of mine lead to neighboring areas were up to 60%. Therefore, Pb isotope ratios can be a good source tracer for areas around abandoned mines.


Assuntos
Monitoramento Ambiental/métodos , Chumbo/análise , Mineração , Poluentes do Solo/análise , Poluição Ambiental/estatística & dados numéricos , Isótopos/análise , República da Coreia
16.
J Microbiol Biotechnol ; 23(11): 1509-18, 2013 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-23966020

RESUMO

An agar-degrading bacterium, designated as strain G7(T), was isolated from a coastal seawater sample from Gaya Island (Gayado in Korean), Republic of Korea. The isolated strain G7(T) is gram-negative, rod shaped, aerobic, non-motile, and non-pigmented. A similarity search based on its 16S rRNA gene sequence revealed that it shares 95.5%, 90.6%, and 90.0% similarity with the 16S rRNA gene sequences of Catenovulum agarivorans YM01(T), Algicola sagamiensis, and Bowmanella pacifica W3-3A(T), respectively. Phylogenetic analyses demonstrated that strain G7(T) formed a distinct monophyletic clade closely related to species of the family Alteromonadaceae in the Alteromonas-like Gammaproteobacteria. The G+C content of strain G7(T) was 41.12 mol%. The DNA-DNA hybridization value between strain G7(T) and the phylogenetically closest strain YM01(T) was 19.63%. The genomes of G7(T) and YM01(T) had an average ANIb value of 70.00%. The predominant isoprenoid quinone of this particular strain was ubiquinone-8, whereas that of C. agarivorans YM01(T) was menaquinone-7. The major fatty acids of strain G7(T) were Iso-C15:0(41.47%), Anteiso-C15:0(22.99%), and C16:1ω7c/iso-C15:02-OH (8.85%), which were quite different from those of YM01(T). Comparison of the phenotypic characteristics related to carbon utilization, enzyme production, and susceptibility to antibiotics also demonstrated that strain G7(T) is distinct from C. agarivorans YM01T. Based on its phenotypic, chemotaxonomic, and phylogenetic distinctiveness, strain G7(T) was considered a novel genus and species in the Gammaproteobacteria, for which the name Gayadomonas joobiniege gen. nov. sp. nov. (ATCC BAA-2321 = DSM25250(T) = KCTC23721(T)) is proposed.


Assuntos
Ágar/metabolismo , Alteromonadaceae/classificação , Alteromonadaceae/metabolismo , Água do Mar/microbiologia , Alteromonadaceae/genética , Alteromonadaceae/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácidos Graxos/análise , Hidrólise , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Transmissão , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Filogenia , Quinonas/análise , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA
17.
Water Res ; 47(3): 1136-46, 2013 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-23253471

RESUMO

In an effort to assess the potential use of ZVMg in contaminant treatments, we examined Cr(VI) reduction mediated by ZVMg particles under neutral pH conditions. The reduction of Cr(VI) was tested with batch experiments by varying [Cr(VI)](0) (4.9, 9.6, 49.9 or 96.9 µM) in the presence of 50 mg/L ZVMg particles ([Mg(0)](0) = 2.06 mM) at pH 7 buffered with 50 mM Na-MOPS. When [Cr(VI)](0) = 4.9 or 9.6 µM, Cr(VI) was completely reduced within 60 min. At higher [Cr(VI)](0) (49.9 or 96.9 µM), by contrast, the reduction became retarded at >120 min likely due to rapid ZVMg dissolution in water and surface precipitation of Cr(III) on ZVMg particles. Surface precipitation was observed only when [Cr(VI)](0) = 49.9 or 96.9 µM and increased with increasing [Cr(VI)](0). The effect of dissolved oxygen was negligible on the rate and extent of Cr(VI) reduction. Experimental results indicated that Cr(VI) was reduced not directly by ZVMg but by reactive intermediates produced from ZVMg-water reaction under the experimental conditions employed in this study. In addition, the observed rates of Cr(VI) reduction appeared to follow an order below unity (0.19) with respect to [Cr(VI)](0). These results imply that ZVMg-mediated Cr(VI) reduction likely occurred via an alternative mechanism to the direct surface-mediated reduction typically observed for other zero-valent metals. Rapid and complete Cr(VI) reduction was achieved when a mass ratio of [ZVMg](0):[Cr(VI)](0) ≥ 100 at neutral pH under both oxic and anoxic conditions. Our results highlights the potential for ZVMg to be used in Cr(VI) treatments especially under neutral pH conditions in the presence of dissolved oxygen.


Assuntos
Cromo/química , Magnésio/química , Concentração de Íons de Hidrogênio , Oxigênio/química , Poluentes Químicos da Água/química
18.
J Biol Chem ; 286(17): 15608-17, 2011 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-21402710

RESUMO

Inhibitors of AMPA-type glutamate ion channels are useful as biochemical probes for structure-function studies and as drug candidates for a number of neurological disorders and diseases. Here, we describe the identification of an RNA inhibitor or aptamer by an in vitro evolution approach and a characterization of its mechanism of inhibition on the sites of interaction by equilibrium binding and on the receptor channel opening rate by a laser-pulse photolysis technique. Our results show that the aptamer is a noncompetitive inhibitor that selectively inhibits the GluA2Q(flip) AMPA receptor subunit without any effect on other AMPA receptor subunits or kainate or NMDA receptors. On the GluA2 subunit, this aptamer preferentially inhibits the flip variant. Furthermore, the aptamer preferentially inhibits the closed-channel state of GluA2Q(flip) with a K(I) = 1.5 µM or by ∼15-fold over the open-channel state. The potency and selectivity of this aptamer rival those of small molecule inhibitors. Together, these properties make this aptamer a promising candidate for the development of water-soluble, highly potent, and GluA2 subunit-selective drugs.


Assuntos
Aptâmeros de Nucleotídeos/farmacologia , Receptores de AMPA/antagonistas & inibidores , Sítios de Ligação , Linhagem Celular , Evolução Molecular Direcionada/métodos , Humanos , Cinética , Transição de Fase/efeitos dos fármacos , Subunidades Proteicas
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