RESUMO
Responses to sunlight exposure of the oil-degrading Dietzia cinnamea P4 strain were evaluated by transcriptional levels of SOS genes, photoreactivation and genes involved in tolerance to high levels of reactive oxygen species. The P4 strain was exposed for 1 and 2 h and the magnitude of level changes in the mRNA was evaluated by qPCR. The results described the activation of the SOS system, with the decline of the repressor lexA gene levels and the concomitant increase of recA and uvrAD genes levels. The genes that participate in the photoreactivation process were also responsive to sunlight. The phrB gene encoding deoxyribodipyrimidine photo-lyase had its expression increased after 1-h exposure, while the phytAB genes showed a progressive increase over the studied period. The protective genes against reactive oxygen species, catalases, superoxides, peroxidases, and thioredoxins, had their expression rates detected under the conditions validated in this study. These results show a fast and coordinated response of genes from different DNA repair and tolerance mechanisms employed by strain P4, suggesting a complex concerted protective action against environmental stressors.
Assuntos
Actinobacteria/genética , Actinobacteria/efeitos da radiação , Regulação Bacteriana da Expressão Gênica/efeitos da radiação , Luz Solar , Adaptação Fisiológica , Proteínas de Bactérias/genética , Reparo do DNA/genética , Hidrolases/genética , Oxirredutases/genética , Reação em Cadeia da Polimerase em Tempo RealRESUMO
The oxidative stress response of the highly resistant actinomycete Dietzia cinnamea P4 after treatment with hydrogen peroxide (H2O2) was assessed in order to depict the possible mechanisms underlying its intrinsic high resistance to DNA damaging agents. We used transcriptional profiling to monitor the magnitude and kinetics of changes in the mRNA levels after exposure to different concentrations of H2O2 at 10 min and 1 h following the addition of the stressor. Catalase and superoxide dismutase genes were induced in different ways, according to the condition applied. Moreover, alkyl hydroperoxide reductase ahpCF, thiol peroxidase, thioredoxin and glutathione genes were upregulated in the presence of H2O2. Expression of peroxidase genes was not detected during the experiment. Overall results point to an actinomycete strain endowed with a set of enzymatic defenses against oxidative stress and with the main genes belonging to a functional SOS system (lexA, recA, uvrD), including suppression of lexA repressor, concomitantly to recA and uvrD gene upregulation upon H2O2 challenge.
Assuntos
Actinomycetales/efeitos dos fármacos , Actinomycetales/metabolismo , Peróxido de Hidrogênio/efeitos adversos , Estresse Oxidativo , Resposta SOS em Genética/fisiologia , Actinomycetales/enzimologia , Actinomycetales/genética , Proteínas de Bactérias/genética , Catalase/classificação , Catalase/genética , Dano ao DNA/efeitos dos fármacos , DNA Helicases/genética , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Genes Bacterianos , Glutationa/genética , Cinética , Peroxidases/genética , Peroxirredoxinas/genética , Filogenia , RNA Mensageiro/metabolismo , Recombinases Rec A/genética , Resposta SOS em Genética/genética , Análise de Sequência , Serina Endopeptidases/genética , Superóxido Dismutase/genética , Tiorredoxinas/genética , Fatores de Tempo , Regulação para Cima/efeitos dos fármacosRESUMO
Dietzia sp. 111N12-1, isolated from the seawater of South China Sea, shows strong petroleum hydrocarbons degradation activity. Here, we report the draft sequence of approximately 3.7-Mbp genome of this strain. To the best of our knowledge, this is the first genome sequence of Dietzia strain isolated from the sea. The genome sequence may provide fundamental molecular information on elucidating the metabolic pathway of hydrocarbons degradation in this strain.(AU)
Assuntos
Actinomycetales , Genoma Bacteriano , Análise de Sequência de DNA , Biodegradação Ambiental , China , HidrocarbonetosRESUMO
Abstract Dietzia sp. 111N12-1, isolated from the seawater of South China Sea, shows strong petroleum hydrocarbons degradation activity. Here, we report the draft sequence of approximately 3.7-Mbp genome of this strain. To the best of our knowledge, this is the first genome sequence of Dietzia strain isolated from the sea. The genome sequence may provide fundamental molecular information on elucidating the metabolic pathway of hydrocarbons degradation in this strain.
Assuntos
Água do Mar/microbiologia , Actinomycetales/isolamento & purificação , Actinomycetales/genética , Genoma Bacteriano , Hidrocarbonetos/metabolismo , Filogenia , Biodegradação Ambiental , Actinomycetales/classificação , Actinomycetales/metabolismo , Petróleo/metabolismo , Sequência de Bases , ChinaRESUMO
Dietzia sp. 111N12-1, isolated from the seawater of South China Sea, shows strong petroleum hydrocarbons degradation activity. Here, we report the draft sequence of approximately 3.7-Mbp genome of this strain. To the best of our knowledge, this is the first genome sequence of Dietzia strain isolated from the sea. The genome sequence may provide fundamental molecular information on elucidating the metabolic pathway of hydrocarbons degradation in this strain.
Assuntos
Actinomycetales/genética , Actinomycetales/isolamento & purificação , Genoma Bacteriano , Hidrocarbonetos/metabolismo , Água do Mar/microbiologia , Actinomycetales/classificação , Actinomycetales/metabolismo , Sequência de Bases , Biodegradação Ambiental , China , Petróleo/metabolismo , FilogeniaRESUMO
The interest in production of natural colorants by microbial fermentation has been currently increased. The effects of D-glucose concentration (3.18-36.82 g/L), inoculum size (12.5 × 10(9)-49.5 × 10(9) cfu cells/mL) and air-flow rate (1.95-12.05 L/L min) on the biomass, total carotenoid and canthaxanthin (CTX) accumulation of Dietzia natronolimnaea HS-1 in a batch bioreactor was scrutinized using a response surface methodology-central composite rotatable design (RSM-CCRD). Second-order polynomial models with high R (2) values ranging from 0.978 to 0.990 were developed for the studied responses using multiple linear regression analysis. The models showed the maximum cumulative amounts of biomass (7.85 g/L), total carotenoid (5.48 mg/L) and CTX (4.99 mg/L) could be achieved at 23.38 g/L of D-glucose, 31.2 × 10(9) cfu cells/mL of inoculation intensity and air-flow rate of 7.85 L/L min. The predicted values for optimum conditions were in good agreement with experimental data.
Assuntos
Actinobacteria/crescimento & desenvolvimento , Actinobacteria/metabolismo , Cantaxantina/biossíntese , Aerobiose , Ar , Carga Bacteriana , Técnicas de Cultura Celular por Lotes , Biomassa , Reatores Biológicos/microbiologia , Glucose/metabolismo , Modelos EstatísticosRESUMO
The interest in production of natural colorants by microbial fermentation has been currently increased. The effects of D-glucose concentration (3.18-36.82 g/L), inoculum size (12.5 x 10(9)-49.5 x 10(9) cfu cells/mL) and air-flow rate (1.95-12.05 L/L min) on the biomass, total carotenoid and canthaxanthin (CTX) accumulation of Dietzia natronolimnaea HS-1 in a batch bioreactor was scrutinized using a response surface methodology-central composite rotatable design (RSM-CCRD). Second-order polynomial models with high R² values ranging from 0.978 to 0.990 were developed for the studied responses using multiple linear regression analysis. The models showed the maximum cumulative amounts of biomass (7.85 g/L), total carotenoid (5.48 mg/L) and CTX (4.99 mg/L) could be achieved at 23.38 g/L of D-glucose, 31.2 x 10(9) cfu cells/mL of inoculation intensity and air-flow rate of 7.85 L/L min. The predicted values for optimum conditions were in good agreement with experimental data.
Assuntos
Actinobacteria/crescimento & desenvolvimento , Actinobacteria/metabolismo , Cantaxantina/biossíntese , Aerobiose , Ar , Carga Bacteriana , Técnicas de Cultura Celular por Lotes , Biomassa , Reatores Biológicos/microbiologia , Glucose/metabolismo , Modelos EstatísticosRESUMO
The interest in production of natural colorants by microbial fermentation has been currently increased. The effects of D-glucose concentration (3.18-36.82 g/L), inoculum size (12.5 x 10(9)-49.5 x 10(9) cfu cells/mL) and air-flow rate (1.95-12.05 L/L min) on the biomass, total carotenoid and canthaxanthin (CTX) accumulation of Dietzia natronolimnaea HS-1 in a batch bioreactor was scrutinized using a response surface methodology-central composite rotatable design (RSM-CCRD). Second-order polynomial models with high R² values ranging from 0.978 to 0.990 were developed for the studied responses using multiple linear regression analysis. The models showed the maximum cumulative amounts of biomass (7.85 g/L), total carotenoid (5.48 mg/L) and CTX (4.99 mg/L) could be achieved at 23.38 g/L of D-glucose, 31.2 x 10(9) cfu cells/mL of inoculation intensity and air-flow rate of 7.85 L/L min. The predicted values for optimum conditions were in good agreement with experimental data.
Assuntos
Actinobacteria/crescimento & desenvolvimento , Actinobacteria/metabolismo , Cantaxantina/biossíntese , Aerobiose , Ar , Carga Bacteriana , Técnicas de Cultura Celular por Lotes , Biomassa , Reatores Biológicos/microbiologia , Glucose/metabolismo , Modelos EstatísticosRESUMO
The petroleum-derived degrading Dietzia cinnamea strain P4 recently had its genome sequenced and annotated. This allowed employing the data on genes that are involved in the degradation of n-alkanes. To examine the physiological behavior of strain P4 in the presence of n-alkanes, the strain was grown under varying conditions of pH and temperature. D. cinnamea P4 was able to grow at pH 7.0-9.0 and at temperatures ranging from 35 ºC to 45 ºC. Experiments of gene expression by real-time quantitative RT-PCR throughout the complete growth cycle clearly indicated the induction of the regulatory gene alkU (TetR family) during early growth. During the logarithmic phase, a large increase in transcriptional levels of a lipid transporter gene was noted. Also, the expression of a gene that encodes the protein fused rubredoxin-alkane monooxygenase was enhanced. Both genes are probably under the influence of the AlkU regulator.
Assuntos
Actinomycetales/genética , Actinomycetales/metabolismo , Alcanos/metabolismo , Perfilação da Expressão Gênica , Genes Bacterianos , Hidrocarbonetos/metabolismo , Redes e Vias Metabólicas/genética , Actinomycetales/crescimento & desenvolvimento , Biotransformação , Concentração de Íons de Hidrogênio , Reação em Cadeia da Polimerase em Tempo Real , TemperaturaRESUMO
The petroleum-derived degrading Dietzia cinnamea strain P4 recently had its genome sequenced and annotated. This allowed employing the data on genes that are involved in the degradation of n-alkanes. To examine the physiological behavior of strain P4 in the presence of n-alkanes, the strain was grown under varying conditions of pH and temperature. D. cinnamea P4 was able to grow at pH 7.0-9.0 and at temperatures ranging from 35 ºC to 45 ºC. Experiments of gene expression by real-time quantitative RT-PCR throughout the complete growth cycle clearly indicated the induction of the regulatory gene alkU (TetR family) during early growth. During the logarithmic phase, a large increase in transcriptional levels of a lipid transporter gene was noted. Also, the expression of a gene that encodes the protein fused rubredoxin-alkane monooxygenase was enhanced. Both genes are probably under the influence of the AlkU regulator.(AU)
Assuntos
Indústria de Petróleo e Gás , Hidrocarbonetos , Palmitatos , Biodegradação AmbientalRESUMO
The petroleum-derived degrading Dietzia cinnamea strain P4 recently had its genome sequenced and annotated. This allowed employing the data on genes that are involved in the degradation of n-alkanes. To examine the physiological behavior of strain P4 in the presence of n-alkanes, the strain was grown under varying conditions of pH and temperature. D. cinnamea P4 was able to grow at pH 7.0-9.0 and at temperatures ranging from 35 ºC to 45 ºC. Experiments of gene expression by real-time quantitative RT-PCR throughout the complete growth cycle clearly indicated the induction of the regulatory gene alkU (TetR family) during early growth. During the logarithmic phase, a large increase in transcriptional levels of a lipid transporter gene was noted. Also, the expression of a gene that encodes the protein fused rubredoxin-alkane monooxygenase was enhanced. Both genes are probably under the influence of the AlkU regulator.
Assuntos
Actinomycetales/genética , Actinomycetales/metabolismo , Alcanos/metabolismo , Perfilação da Expressão Gênica , Genes Bacterianos , Hidrocarbonetos/metabolismo , Redes e Vias Metabólicas/genética , Actinomycetales/crescimento & desenvolvimento , Biotransformação , Concentração de Íons de Hidrogênio , Reação em Cadeia da Polimerase em Tempo Real , TemperaturaRESUMO
Foram analisados 24 isolados bacterianos oriundos de leite e pele de búfalas (Bubalus bubalis), os quais foram previamente identificados como Rhodococcus equi com o auxílio de fenotipia concisa. Testes fenotípicos complementares e ferramentas moleculares foram utilizados com o objetivo de caracterizar esses isolados, bem como diferenciá-los de outros microrganismos intimamente relacionados. Observaram-se três fenótipos distintos, porém a identificação dos isolados foi inconclusiva. Apenas um dos isolados foi comprovado como sendo R. equi com a realização da PCR espécie-específica, sequenciamento e análise dos fragmentos de DNA. Os demais isolados só foram identificados pelo sequenciamento de fragmento do gene que codifica a região 16S do rRNA universal de bactérias, indicando tratar-se de Dietzia maris. O perfil de susceptibilidade aos antimicrobianos revelou maior resistência dos isolados de D. maris para oxacilina (96 por cento) e rifampicina (87 por cento). O isolado de R. equi apresentou resistência à amicacina, oxacilina, penicilina, rifampicina e tetraciclina. Alerta-se para o risco da incorreta identificação dos isolados baseados em testes fenotípicos concisos e para a necessidade de utilização de testes complementares para diferenciação entre R. equi e D. maris.(AU)
Twenty-four bacterial isolates from milk and skin of buffalo females (Bubalus bubalis), which previously had been identified as Rhodococcus equi by using a restricted number of phenotypical tests for bacterial characterization, were analyzed. The goal of this study was to perform the characterization of these isolates, as well as the differentiation of other microorganisms closely related by using additional phenotypical tests and molecular tools. Based on the phenotypical results, three different biotypes were obtained. However, the identification of the isolates was inconclusive. Only one of the isolates was confirmed as R. equi by the PCR specifically for this species, as well DNA sequencing and DNA fragment analysis. All the other isolates only could be precisely identified after the DNA sequencing, and they were characterized as Dietzia maris. The sensitivity profile to antimicrobials demonstrated the highest resistance of D. maris to oxacillin and rifampin, 96 percent and 87 percent, respectively. R. equi isolate, presented resistance to amikacin, oxacillin, penicillin, rifampin, and tetracycline. Thus, it is important to alert for the risk of the incorrect identification of the bacterial isolates by using diagnostic analysis based on phenotypical tests in order to differentiate R. equi and D. maris, besides the necessity to use complementary tests for differentiation of these microorganisms.(AU)
Assuntos
Animais , Rhodococcus equi/isolamento & purificação , Corynebacterium/classificação , Corynebacterium/isolamento & purificação , Análise de Sequência de DNA/métodos , Análise de Sequência de DNA/veterinária , Búfalos/genéticaRESUMO
Foram analisados 24 isolados bacterianos oriundos de leite e pele de búfalas (Bubalus bubalis), os quais foram previamente identificados como Rhodococcus equi com o auxílio de fenotipia concisa. Testes fenotípicos complementares e ferramentas moleculares foram utilizados com o objetivo de caracterizar esses isolados, bem como diferenciá-los de outros microrganismos intimamente relacionados. Observaram-se três fenótipos distintos, porém a identificação dos isolados foi inconclusiva. Apenas um dos isolados foi comprovado como sendo R. equi com a realização da PCR espécie-específica, sequenciamento e análise dos fragmentos de DNA. Os demais isolados só foram identificados pelo sequenciamento de fragmento do gene que codifica a região 16S do rRNA universal de bactérias, indicando tratar-se de Dietzia maris. O perfil de susceptibilidade aos antimicrobianos revelou maior resistência dos isolados de D. maris para oxacilina (96 por cento) e rifampicina (87 por cento). O isolado de R. equi apresentou resistência à amicacina, oxacilina, penicilina, rifampicina e tetraciclina. Alerta-se para o risco da incorreta identificação dos isolados baseados em testes fenotípicos concisos e para a necessidade de utilização de testes complementares para diferenciação entre R. equi e D. maris.
Twenty-four bacterial isolates from milk and skin of buffalo females (Bubalus bubalis), which previously had been identified as Rhodococcus equi by using a restricted number of phenotypical tests for bacterial characterization, were analyzed. The goal of this study was to perform the characterization of these isolates, as well as the differentiation of other microorganisms closely related by using additional phenotypical tests and molecular tools. Based on the phenotypical results, three different biotypes were obtained. However, the identification of the isolates was inconclusive. Only one of the isolates was confirmed as R. equi by the PCR specifically for this species, as well DNA sequencing and DNA fragment analysis. All the other isolates only could be precisely identified after the DNA sequencing, and they were characterized as Dietzia maris. The sensitivity profile to antimicrobials demonstrated the highest resistance of D. maris to oxacillin and rifampin, 96 percent and 87 percent, respectively. R. equi isolate, presented resistance to amikacin, oxacillin, penicillin, rifampin, and tetracycline. Thus, it is important to alert for the risk of the incorrect identification of the bacterial isolates by using diagnostic analysis based on phenotypical tests in order to differentiate R. equi and D. maris, besides the necessity to use complementary tests for differentiation of these microorganisms.