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  1 / 103747 MEDLINE  
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[PMID]:29385201
[Au] Autor:Zhou Z; Liu F; Zhang X; Zhou X; Zhong Z; Su H; Li J; Li H; Feng F; Lan J; Zhang Z; Fu H; Hu Y; Cao S; Chen W; Deng J; Yu J; Zhang W; Peng G
[Ad] Endereço:The Key Laboratory of Animal Disease and Human Health of Sichuan Province, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China.
[Ti] Título:Cellulose-dependent expression and antibacterial characteristics of surfactin from Bacillus subtilis HH2 isolated from the giant panda.
[So] Source:PLoS One;13(1):e0191991, 2018.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Surfactin secreted by Bacillus subtilis can confer strong, diverse antipathogenic effects, thereby benefitting the host. Carbon source is an important factor for surfactin production. However, the mechanism that bacteria utilize cellulose, the most abundant substance in the intestines of herbivores, to produce surfactin remains unclear. Here, we used B. subtilis HH2, isolated from the feces of a giant panda, as a model to determine changes in surfactin expression in the presence of different concentrations of cellulose by quantitative polymerase chain reaction and high-performance liquid chromatography. We further investigated the antimicrobial effects of surfactin against three common intestinal pathogens (Escherichia coli, Staphylococcus aureus, and Salmonella enterica) and its resistance to high temperature (60-121°C), pH (1-12), trypsin (100-300 µg/mL, pH 8), and pepsin (100-300 µg/mL, pH 2). The results showed that the surfactin expressed lowest in bacteria cultured in the presence of 1% glucose medium as the carbon source, whereas increased in an appropriate cellulose concentration (0.67% glucose and 0.33% cellulose). The surfactin could inhibit E. coli and Staphylococcus aureus, but did not affect efficiently for Salmonella enterica. The antibacterial ability of surfactin did not differ according to temperature (60-100°C), pH (2-11), trypsin (100-300 µg/mL), and pepsin (100-300 µg/mL; P > 0.05), but decreased significantly at extreme environments (121°C, pH 1 or 12; P < 0.05) compared with that in the control group (37°C, pH = 7, without any protease). In conclusion, our findings indicated that B. subtilis HH2 could increase surfactin expression in an appropriate cellulose environment and thus provide benefits to improve the intestinal health of herbivores.
[Mh] Termos MeSH primário: Antibacterianos/metabolismo
Bacillus subtilis/metabolismo
Celulose/metabolismo
Lipopeptídeos/metabolismo
[Mh] Termos MeSH secundário: Animais
Antibacterianos/farmacologia
Meios de Cultura
Lipopeptídeos/farmacologia
Ursidae
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Anti-Bacterial Agents); 0 (Culture Media); 0 (Lipopeptides); 9004-34-6 (Cellulose)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180309
[Lr] Data última revisão:
180309
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180201
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0191991


  2 / 103747 MEDLINE  
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[PMID]:29377912
[Au] Autor:Robertson MJ; Soibam B; O'Leary JG; Sampaio LC; Taylor DA
[Ad] Endereço:Scientific Stem Cell, Texas Heart Institute, Houston, Texas, United States of America.
[Ti] Título:Recellularization of rat liver: An in vitro model for assessing human drug metabolism and liver biology.
[So] Source:PLoS One;13(1):e0191892, 2018.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Liver-like organoids that recapitulate the complex functions of the whole liver by combining cells, scaffolds, and mechanical or chemical cues are becoming important models for studying liver biology and drug metabolism. The advantages of growing cells in three-dimensional constructs include enhanced cell-cell and cell-extracellular matrix interactions and preserved cellular phenotype including, prevention of de-differentiation. In the current study, biomimetic liver constructs were made via perfusion decellularization of rat liver, with the goal of maintaining the native composition and structure of the extracellular matrix. We optimized our decellularization process to produce liver scaffolds in which immunogenic residual DNA was removed but glycosaminoglycans were maintained. When the constructs were recellularized with rat or human liver cells, the cells remained viable, capable of proliferation, and functional for 28 days. Specifically, the cells continued to express cytochrome P450 genes and maintained their ability to metabolize a model drug, midazolam. Microarray analysis showed an upregulation of genes involved in liver regeneration and fibrosis. In conclusion, these liver constructs have the potential to be used as test beds for studying liver biology and drug metabolism.
[Mh] Termos MeSH primário: Fígado/citologia
Modelos Animais
Farmacocinética
[Mh] Termos MeSH secundário: Animais
Reatores Biológicos
Adesão Celular
Proliferação Celular
Meios de Cultura
Matriz Extracelular
Hepatócitos/efeitos dos fármacos
Hepatócitos/metabolismo
Seres Humanos
Técnicas In Vitro
Ratos
Ratos Sprague-Dawley
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Culture Media)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180309
[Lr] Data última revisão:
180309
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180130
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0191892


  3 / 103747 MEDLINE  
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[PMID]:29267658
[Au] Autor:Ferrúa CP; Centeno EGZ; Rosa LCD; Amaral CCD; Severo RF; Sarkis-Onofre R; Nascimento GG; Cordenonzi G; Bast RK; Demarco FF; Nedel F
[Ad] Endereço:Universidade Católica de Pelotas - UCPel, Program in Health and Behavior, Pelotas, RS, Brazil.
[Ti] Título:How has dental pulp stem cells isolation been conducted? A scoping review.
[So] Source:Braz Oral Res;31:e87, 2017 Dec 18.
[Is] ISSN:1807-3107
[Cp] País de publicação:Brazil
[La] Idioma:eng
[Ab] Resumo:The objective of this study was to realize a scoping review the literature in order to identify the profile of DPSCs isolation and analyze the possible risk factors that could change the native behavior of these cells. An initial search was conducted using the following MeSH terms: "(dental pulp stem cell [MeSH])"; "(dental pulp [MeSH])" AND "(stem cell [MeSH])"; "("dental pulp stem cell" [MeSH]")". The electronic search was done without date restriction up to and including April 2014, in PubMed, Scopus, Scielo and ISI Web of Knowledge databases. Studies were submitted to inclusion and exclusion criteria and 222 articles were included. Data showed that over the past 15 years many studies have been conducted using DPSCs. However this is the first systematic review regarding the isolation of stem cell, and more specifically of dental pulp stem cells. The isolation of dental pulp stem cells showed great variability, hampering the development of standard protocols to achieve in vitro dental pulp stem cells with similar characteristics. This scoping review combined, for the first time, the methodologies used for dental pulp stem isolation, highlighting the most frequently used.
[Mh] Termos MeSH primário: Polpa Dentária/citologia
Células-Tronco/citologia
[Mh] Termos MeSH secundário: Técnicas de Cultura de Células
Colagenases
Meios de Cultura
Seres Humanos
Viés de Publicação
Fatores de Risco
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Nm] Nome de substância:
0 (Culture Media); EC 3.4.24.- (Collagenases)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180308
[Lr] Data última revisão:
180308
[Sb] Subgrupo de revista:D; IM
[Da] Data de entrada para processamento:171222
[St] Status:MEDLINE


  4 / 103747 MEDLINE  
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[PMID]:29267499
[Au] Autor:Santos APP; Silva MDS; Costa EVL; Rufino RD; Santos VA; Ramos CS; Sarubbo LA; Porto ALF
[Ad] Endereço:Departamento de Morfologia e Fisiologia Animal, Universidade Federal Rural de Pernambuco, Recife, PE, Brasil.
[Ti] Título:Production and characterization of a biosurfactant produced by Streptomyces sp. DPUA 1559 isolated from lichens of the Amazon region.
[So] Source:Braz J Med Biol Res;51(2):e6657, 2017 Dec 11.
[Is] ISSN:1414-431X
[Cp] País de publicação:Brazil
[La] Idioma:eng
[Ab] Resumo:Surfactants are amphipathic compounds containing both hydrophilic and hydrophobic groups, capable to lower the surface or interfacial tension. Considering the advantages of the use of biosurfactants produced by microorganisms, the aim of this paper was to develop and characterize a biosurfactant produced by Streptomyces sp. DPUA1559 isolated from lichens of the Amazon region. The microorganism was cultured in a mineral medium containing 1% residual frying soybean oil as the carbon source. The kinetics of biosurfactant production was accompanied by reducing the surface tension of the culture medium from 60 to values around 27.14 mN/m, and by the emulsification index, which showed the efficiency of the biosurfactant as an emulsifier of hydrophobic compounds. The yield of the isolated biosurfactant was 1.74 g/L, in addition to the excellent capability of reducing the surface tension (25.34 mN/m), as observed from the central composite rotational design when the biosurfactant was produced at pH 8.5 at 28°C. The critical micelle concentration of the biosurfactant was determined as 0.01 g/mL. The biosurfactant showed thermal and pH stability regarding the surface tension reduction, and tolerance under high salt concentrations. The isolated biosurfactant showed no toxicity to the micro-crustacean Artemia salina, and to the seeds of lettuce (Lactuca sativa L.) and cabbage (Brassica oleracea L.). The biochemistry characterization of the biosurfactant showed a single protein band, an acid character and a molecular weight around 14.3 kDa, suggesting its glycoproteic nature. The results are promising for the industrial application of this new biosurfactant.
[Mh] Termos MeSH primário: Líquens/microbiologia
Streptomyces/metabolismo
Tensoativos/metabolismo
[Mh] Termos MeSH secundário: Análise de Variância
Contagem de Colônia Microbiana
Meios de Cultura
Eletroforese em Gel de Poliacrilamida
Fermentação
Concentração de Íons de Hidrogênio
Valores de Referência
Sementes/efeitos dos fármacos
Óleo de Soja/química
Espectroscopia de Infravermelho com Transformada de Fourier
Streptomyces/crescimento & desenvolvimento
Streptomyces/isolamento & purificação
Tensão Superficial
Tensoativos/análise
Tensoativos/química
Temperatura Ambiente
Fatores de Tempo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Culture Media); 0 (Surface-Active Agents); 8001-22-7 (Soybean Oil)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180308
[Lr] Data última revisão:
180308
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171222
[St] Status:MEDLINE


  5 / 103747 MEDLINE  
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[PMID]:28463419
[Au] Autor:Lin C; Khetani SR
[Ad] Endereço:School of Biomedical Engineering, Colorado State University, Fort Collins, Colorado.
[Ti] Título:Micropatterned Co-Cultures of Human Hepatocytes and Stromal Cells for the Assessment of Drug Clearance and Drug-Drug Interactions.
[So] Source:Curr Protoc Toxicol;72:14.17.1-14.17.23, 2017 May 02.
[Is] ISSN:1934-9262
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Drug clearance rates from the body can determine drug exposure that can affect efficacy or toxicity. Thus, accurate prediction of drug clearance during preclinical development can help guide dose selection in humans, but animal testing is not always predictive of human outcomes. Because hepatic drug metabolism is a rate-limiting step in the overall clearance of many drugs, primary human hepatocytes (PHHs) in suspension cultures or monolayers are used for drug clearance predictions. Yet, the precipitous decline in drug metabolism capacity can lead to significant underestimation of clearance rates, particularly for low turnover compounds that have desirable one-pill-a-day dosing regimens. In contrast, micropatterned co-cultures (MPCCs) of PHHs and fibroblasts display phenotypic stability for several weeks and can help mitigate the limitations of conventional cultures. Here, we describe protocols to create and use MPCCs for drug clearance predictions, and for modeling clinically-relevant drug-drug interactions that can affect drug clearance. © 2017 by John Wiley & Sons, Inc.
[Mh] Termos MeSH primário: Técnicas de Cocultura/métodos
Interações Medicamentosas
Hepatócitos/metabolismo
Preparações Farmacêuticas/metabolismo
Células Estromais/metabolismo
[Mh] Termos MeSH secundário: Células 3T3
Animais
Técnicas de Cultura de Células
Meios de Cultura
Fibroblastos/metabolismo
Hepatócitos/ultraestrutura
Seres Humanos
Taxa de Depuração Metabólica
Camundongos
Fenótipo
Células Estromais/ultraestrutura
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Culture Media); 0 (Pharmaceutical Preparations)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180308
[Lr] Data última revisão:
180308
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170503
[St] Status:MEDLINE
[do] DOI:10.1002/cptx.23


  6 / 103747 MEDLINE  
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[PMID]:28463418
[Au] Autor:Liu C; Sekine S; Song B; Ito K
[Ad] Endereço:Laboratory of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan.
[Ti] Título:Use of Primary Rat Hepatocytes for Prediction of Drug-Induced Mitochondrial Dysfunction.
[So] Source:Curr Protoc Toxicol;72:14.16.1-14.16.10, 2017 May 02.
[Is] ISSN:1934-9262
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Mitochondrial dysfunction plays a central role in drug-induced liver injury. To evaluate drug-induced mitochondrial impairment, several isolated mitochondria- or cell line-based assays have been reported. Among them, culturing HepG2 cells in galactose provides a remarkable method to assess mitochondrial toxicity by activating mitochondrial aerobic respiration. We applied this assay to primary rat hepatocytes by culturing cells in galactose and hyperoxia to enhance the evaluation of metabolism-related drug-induced mitochondrial toxicity. Conventional culture of primary hepatocytes under high-glucose and hypoxic conditions could force cells to switch energy generation to glycolysis. By contrast, cells cultured in galactose and hyperoxia could maintain energy generation from mitochondrial aerobic respiration, which is consistent with physiological conditions, and consequently improve the susceptibility of cells to mitochondrial toxicants. Measuring the toxicities of test compounds in primary rat hepatocytes cultured in modified conditions provides a useful model to identify mitochondrial dysfunction-mediated drug-induced hepatotoxicity. © 2017 by John Wiley & Sons, Inc.
[Mh] Termos MeSH primário: Hepatócitos/efeitos dos fármacos
Doenças Mitocondriais/induzido quimicamente
Doenças Mitocondriais/patologia
[Mh] Termos MeSH secundário: Animais
Respiração Celular
Doença Hepática Induzida por Substâncias e Drogas
Meios de Cultura
Metabolismo Energético
Previsões
Galactose/metabolismo
Hepatócitos/enzimologia
Hiperóxia/metabolismo
L-Lactato Desidrogenase/análise
Consumo de Oxigênio
Cultura Primária de Células
Ratos
Ratos Sprague-Dawley
Testes de Toxicidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Culture Media); EC 1.1.1.27 (L-Lactate Dehydrogenase); X2RN3Q8DNE (Galactose)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180308
[Lr] Data última revisão:
180308
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170503
[St] Status:MEDLINE
[do] DOI:10.1002/cptx.24


  7 / 103747 MEDLINE  
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[PMID]:28463416
[Au] Autor:Kaja S; Payne AJ; Naumchuk Y; Koulen P
[Ad] Endereço:Departments of Ophthalmology and Molecular Pharmacology & Therapeutics, Stritch School of Medicine, Loyola University Chicago, Maywood, Illinois.
[Ti] Título:Quantification of Lactate Dehydrogenase for Cell Viability Testing Using Cell Lines and Primary Cultured Astrocytes.
[So] Source:Curr Protoc Toxicol;72:2.26.1-2.26.10, 2017 May 02.
[Is] ISSN:1934-9262
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Drug discovery heavily relies on cell viability studies to assess the potential toxicity of drug candidates. L-Lactate dehydrogenase (LDH) is a cytoplasmic enzyme that catalyzes the concomitant interconversions of pyruvate to L-lactate and NADH to NAD during glycolysis, and the reverse reactions during the Cori cycle. In response to cellular damage, induced by endogenous cellular mechanisms or as a result of exogenously applied insults, LDH is released from the cytoplasm into the extracellular environment. Its stability in cell culture medium makes it a well-suited correlate for the presence of damage and toxicity in tissues and cells. We herein present protocols for a reproducible and validated LDH assay optimized for several cell types. In contrast to commercially available LDH assays, often associated with proprietary formulations and high cost, our protocols provide ample opportunities for experiment-specific optimization with low variability and cost. © 2017 by John Wiley & Sons, Inc.
[Mh] Termos MeSH primário: Astrócitos/enzimologia
Sobrevivência Celular
L-Lactato Desidrogenase/análise
Toxicologia/métodos
[Mh] Termos MeSH secundário: Animais
Técnicas de Cultura de Células
Linhagem Celular
Linhagem Celular Tumoral
Meios de Cultura/química
Citoplasma/enzimologia
Espaço Extracelular/enzimologia
Glicólise
Seres Humanos
Neurônios/efeitos dos fármacos
Fármacos Neuroprotetores/farmacologia
Nervo Óptico/citologia
Nervo Óptico/efeitos dos fármacos
Cultura Primária de Células
Ratos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Culture Media); 0 (Neuroprotective Agents); EC 1.1.1.27 (L-Lactate Dehydrogenase)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180308
[Lr] Data última revisão:
180308
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170503
[St] Status:MEDLINE
[do] DOI:10.1002/cptx.21


  8 / 103747 MEDLINE  
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[PMID]:29412221
[Au] Autor:Daniela Ferreira Araújo B; Luciana Oliveira P; Izabel Cristina Rodrigues da S; Ricardo Bentes A; Ana Cristina Barreto B
[Ad] Endereço:Universidade de Brasília - UnB, Institute of Biological Sciences, Laboratory of Nanobiotechnology, Brasília, DF, Brazil.
[Ti] Título:Culture of human dental pulp cells at variable times post-tooth extraction.
[So] Source:Braz Oral Res;32:e003, 2018 Feb 01.
[Is] ISSN:1807-3107
[Cp] País de publicação:Brazil
[La] Idioma:eng
[Ab] Resumo:The aim of this study was to investigate the viability of human dental pulp cells from extracted teeth kept at standard room temperature and atmospheric pressure for different periods of time. Twenty-one healthy permanent teeth were used. They were divided into five groups according to the expected time from extraction to processing. One group was tested immediately after extraction; the other groups were each tested at one of the following time points: 30 minutes, 1 hour, 2 hours, and 5 hours post-extraction. Cell morphology was analysed by light microscopy; cell proliferation was analysed using MTT assay and by counting the viable cells in a haemocytometer. Similar results were observed in all groups (p < 0.05). A delay of up to five hours for tooth processing and tissue collection does not preclude the establishment of dental pulp cell cultures, affect the morphology of these cells, or reduce their proliferative potential.
[Mh] Termos MeSH primário: Técnicas de Cultura de Células/métodos
Polpa Dentária/citologia
Extração Dentária
[Mh] Termos MeSH secundário: Análise de Variância
Contagem de Células
Proliferação Celular
Sobrevivência Celular
Células Cultivadas
Meios de Cultura
Seres Humanos
Reprodutibilidade dos Testes
Sais de Tetrazólio
Tiazóis
Fatores de Tempo
Preservação de Tecido/métodos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Culture Media); 0 (Tetrazolium Salts); 0 (Thiazoles); EUY85H477I (thiazolyl blue)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180307
[Lr] Data última revisão:
180307
[Sb] Subgrupo de revista:D; IM
[Da] Data de entrada para processamento:180208
[St] Status:MEDLINE


  9 / 103747 MEDLINE  
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[PMID]:29339722
[Au] Autor:Sousa DZ; Visser M; van Gelder AH; Boeren S; Pieterse MM; Pinkse MWH; Verhaert PDEM; Vogt C; Franke S; Kümmel S; Stams AJM
[Ad] Endereço:Laboratory of Microbiology, Wageningen University & Research, Stippeneng 4, 6708 WE, Wageningen, The Netherlands.
[Ti] Título:The deep-subsurface sulfate reducer Desulfotomaculum kuznetsovii employs two methanol-degrading pathways.
[So] Source:Nat Commun;9(1):239, 2018 01 16.
[Is] ISSN:2041-1723
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Methanol is generally metabolized through a pathway initiated by a cobalamine-containing methanol methyltransferase by anaerobic methylotrophs (such as methanogens and acetogens), or through oxidation to formaldehyde using a methanol dehydrogenase by aerobes. Methanol is an important substrate in deep-subsurface environments, where thermophilic sulfate-reducing bacteria of the genus Desulfotomaculum have key roles. Here, we study the methanol metabolism of Desulfotomaculum kuznetsovii strain 17 , isolated from a 3000-m deep geothermal water reservoir. We use proteomics to analyze cells grown with methanol and sulfate in the presence and absence of cobalt and vitamin B12. The results indicate the presence of two methanol-degrading pathways in D. kuznetsovii, a cobalt-dependent methanol methyltransferase and a cobalt-independent methanol dehydrogenase, which is further confirmed by stable isotope fractionation. This is the first report of a microorganism utilizing two distinct methanol conversion pathways. We hypothesize that this gives D. kuznetsovii a competitive advantage in its natural environment.
[Mh] Termos MeSH primário: Álcool Desidrogenase/metabolismo
Proteínas de Bactérias/metabolismo
Desulfotomaculum/enzimologia
Redes e Vias Metabólicas/genética
Metanol/metabolismo
Metiltransferases/metabolismo
[Mh] Termos MeSH secundário: Álcool Desidrogenase/genética
Proteínas de Bactérias/genética
Cobalto/metabolismo
Cobalto/farmacologia
Meios de Cultura/química
Desulfotomaculum/genética
Expressão Gênica
Perfilação da Expressão Gênica
Hidrólise
Metiltransferases/genética
Oxirredução
Filogenia
Proteômica/métodos
Vitamina B 12/metabolismo
Vitamina B 12/farmacologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (Culture Media); 3G0H8C9362 (Cobalt); EC 1.1.1.1 (Alcohol Dehydrogenase); EC 2.1.1.- (Methyltransferases); EVS87XF13W (cobaltous chloride); P6YC3EG204 (Vitamin B 12); Y4S76JWI15 (Methanol)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180307
[Lr] Data última revisão:
180307
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180118
[St] Status:MEDLINE
[do] DOI:10.1038/s41467-017-02518-9


  10 / 103747 MEDLINE  
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[PMID]:28461451
[Au] Autor:Li L; Jiang W; Lu Y
[Ad] Endereço:Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
[Ti] Título:A Novel Two-Component System, GluR-GluK, Involved in Glutamate Sensing and Uptake in Streptomyces coelicolor.
[So] Source:J Bacteriol;199(18), 2017 09 15.
[Is] ISSN:1098-5530
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Two-component systems (TCSs), the predominant signal transduction pathways employed by bacteria, play important roles in physiological metabolism in Here, a novel TCS, GluR-GluK (encoded by ), which is located divergently from the operon encoding a glutamate uptake system, was identified as being involved in glutamate sensing and uptake as well as antibiotic biosynthesis in Under the condition of minimal medium (MM) supplemented with different concentrations of glutamate, deletion of the operon ( ) resulted in enhanced actinorhodin (ACT) but reduced undecylprodigiosin (RED) and yellow type I polyketide (yCPK) production, suggesting that GluR-GluK plays a differential role in antibiotic biosynthesis. Furthermore, we found that the response regulator GluR directly promotes the expression of under the culture condition of MM with a high concentration of glutamate (75 mM). Using the biolayer interferometry assay, we demonstrated that glutamate acts as the direct signal of the histidine kinase GluK. It was therefore suggested that upon sensing high concentrations of glutamate, GluR-GluK would be activated and thereby facilitate glutamate uptake by increasing expression. Finally, we demonstrated that the role of GluR-GluK in antibiotic biosynthesis is independent of its function in glutamate uptake. Considering the wide distribution of the glutamate-sensing (GluR-GluK) and uptake (GluABCD) module in actinobacteria, it could be concluded that the GluR-GluK signal transduction pathway involved in secondary metabolism and glutamate uptake should be highly conserved in this bacterial phylum. In this study, a novel two-component system (TCS), GluR-GluK, was identified to be involved in glutamate sensing and uptake as well as antibiotic biosynthesis in A possible GluR-GluK working model was proposed. Upon sensing high glutamate concentrations (such as 75 mM), activated GluR-GluK could regulate both glutamate uptake and antibiotic biosynthesis. However, under a culture condition of MM supplemented with low concentrations of glutamate (such as 10 mM), although GluR-GluK is activated, its activity is sufficient only for the regulation of antibiotic biosynthesis. To the best of our knowledge, this is the first report describing a TCS signal transduction pathway for glutamate sensing and uptake in actinobacteria.
[Mh] Termos MeSH primário: Ácido Glutâmico/metabolismo
Histidina Quinase/metabolismo
Transdução de Sinais
Streptomyces coelicolor/metabolismo
Fatores de Transcrição/metabolismo
[Mh] Termos MeSH secundário: Transporte Biológico
Meios de Cultura/química
Deleção de Genes
Regulação da Expressão Gênica
Histidina Quinase/genética
Proteínas de Membrana Transportadoras/genética
Proteínas de Membrana Transportadoras/metabolismo
Óperon
Streptomyces coelicolor/genética
Fatores de Transcrição/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, U.S. GOV'T, NON-P.H.S.; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Culture Media); 0 (Membrane Transport Proteins); 0 (Transcription Factors); 3KX376GY7L (Glutamic Acid); EC 2.7.13.1 (Histidine Kinase)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:180306
[Lr] Data última revisão:
180306
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170503
[St] Status:MEDLINE



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