Seleção de microrganismos endofíticos com potencialidades para a biorremediação de ambientes contaminados com hidrocarbonetos de petróleo e/ou derivados

Detalhes bibliográficos
Autor(a) principal: OLIVEIRA, Natalia Carvalhaes de
Data de Publicação: 2009
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFG
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tde/1795
Resumo: Endophytic microorganisms live inside plants showing no apparently damage for the host, often assisting in survival of plants, helping its growth with production of phytohormones, phosphates solubilization, nitrogen fixation and enzymes production, or they can metabolize organic contaminants, like petroleum and derivates. This work aimed to isolated and identified endophytic microorganisms of plants present in impacted areas, as well as test their ability in petroleum and its derivatives degradation, identify bacteriocin production, to test their nitrogen fixation capability, phosphate solubilization, indol-acetic acid (IAA) and enzymes production. Plant samples were collected, in an area impacted with asphaltic and mud, were superficially disinfected using 70% ethanol, sodium hypochlorite and sterile distilled water. After macerated and fragmented, the samples were incubated at 30°C for about 72 hours, when growth of microorganisms was observed in culture media: Nutrient Agar, TSA (Tryptone Soya Agar) and King medium. The verification of petroleum and derivatives degradation capacity was performed in ELISA plates, exposing the bacteria to a solution of Minimal Medium, the dye DCPIP solution (2,6-dicloroindofenol sodium salt) and petroleum or derivative tested (burning oil, lubricating oil, diesel oil and gasoline). A positive reading for degradation was observed by discoloration of DCPIP. Among nine bacteria tested, three showed degradative activity in different fractions of petroleum, diesel oil and gasoline, and others showed different profiles. This result shows the potential of these microorganisms for application in bioremediation processes. The production of IAA was tested on TSA medium supplemented with tryptophan and only one sample was positive, while none was considered phosphate solubilizing. Testing the nitrogen fixing, samples were inoculated for a free nitrogen culture media, showing growth after 24 hours of incubation (positive results), however it s isolated did not show positive results in the acetylene reduction test, which confirms activity of the enzyme nitrogenase. The samples showed different profiles for the production and activity of bacteriocins against the clinical isolates tested. There were no detected activity of endoglucanase, cellulolytic and pectinolytic enzymes, but esterases, ß-glucosidases, proteases and amylases were positive. Some isolates were identified from the sequencing of 16S DNA, whose sequence was compared to database (GenBank), and then diagnosed as Bacillus cereus, Staphylococcus pasteuri and Pseudomonas sp. The endophytic bacteria isolated from Cerrado plants confirmed their potential for use in bioremediation, plant growth promotion and diverse enzyme production, with potential for use in pharmaceutical, food, textiles, among others.
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spelling VIEIRA, José Daniel Gonçalveshttp://lattes.cnpq.br/1742731776579730http://lattes.cnpq.br/7560948180074168OLIVEIRA, Natalia Carvalhaes de2014-07-29T15:30:35Z2011-05-022009-03-16OLIVEIRA, Natalia Carvalhaes de. Selection of endophytic microorganisms for biorremediation in impacted soils with petroleum hydrocarbons and derivates. 2009. 96 f. Dissertação (Mestrado em Medicina) - Universidade Federal de Goiás, Goiânia, 2009.http://repositorio.bc.ufg.br/tede/handle/tde/1795ark:/38995/0013000004k2fEndophytic microorganisms live inside plants showing no apparently damage for the host, often assisting in survival of plants, helping its growth with production of phytohormones, phosphates solubilization, nitrogen fixation and enzymes production, or they can metabolize organic contaminants, like petroleum and derivates. This work aimed to isolated and identified endophytic microorganisms of plants present in impacted areas, as well as test their ability in petroleum and its derivatives degradation, identify bacteriocin production, to test their nitrogen fixation capability, phosphate solubilization, indol-acetic acid (IAA) and enzymes production. Plant samples were collected, in an area impacted with asphaltic and mud, were superficially disinfected using 70% ethanol, sodium hypochlorite and sterile distilled water. After macerated and fragmented, the samples were incubated at 30°C for about 72 hours, when growth of microorganisms was observed in culture media: Nutrient Agar, TSA (Tryptone Soya Agar) and King medium. The verification of petroleum and derivatives degradation capacity was performed in ELISA plates, exposing the bacteria to a solution of Minimal Medium, the dye DCPIP solution (2,6-dicloroindofenol sodium salt) and petroleum or derivative tested (burning oil, lubricating oil, diesel oil and gasoline). A positive reading for degradation was observed by discoloration of DCPIP. Among nine bacteria tested, three showed degradative activity in different fractions of petroleum, diesel oil and gasoline, and others showed different profiles. This result shows the potential of these microorganisms for application in bioremediation processes. The production of IAA was tested on TSA medium supplemented with tryptophan and only one sample was positive, while none was considered phosphate solubilizing. Testing the nitrogen fixing, samples were inoculated for a free nitrogen culture media, showing growth after 24 hours of incubation (positive results), however it s isolated did not show positive results in the acetylene reduction test, which confirms activity of the enzyme nitrogenase. The samples showed different profiles for the production and activity of bacteriocins against the clinical isolates tested. There were no detected activity of endoglucanase, cellulolytic and pectinolytic enzymes, but esterases, ß-glucosidases, proteases and amylases were positive. Some isolates were identified from the sequencing of 16S DNA, whose sequence was compared to database (GenBank), and then diagnosed as Bacillus cereus, Staphylococcus pasteuri and Pseudomonas sp. The endophytic bacteria isolated from Cerrado plants confirmed their potential for use in bioremediation, plant growth promotion and diverse enzyme production, with potential for use in pharmaceutical, food, textiles, among others.Microrganismos endofíticos vivem no interior de plantas sem causar danos aparentes no hospedeiro, muitas vezes auxiliando na sobrevivência do vegetal, auxiliando seu crescimento com produção de fitormônios, solubilização de fosfatos, fixação de nitrogênio e produção de enzimas, ou metabolizando alguns compostos contaminantes orgânicos, como petróleo e/ou derivados. O objetivo deste trabalho foi isolar e identificar microrganismos endofíticos de vegetais presentes em áreas impactadas, assim como testar sua capacidade de degradação de petróleo e alguns derivados, identificar seu perfil para a produção de bacteriocinas, capacidade de fixação de nitrogênio, solubilização de fosfatos, produção de ácido indol-acético (AIA) e enzimas. Amostras vegetais foram coletadas em uma área impactada com lama asfáltica e desinfectadas superficialmente utilizando etanol 70%, hipoclorito de sódio e água destilada esterilizada. Depois de fragmentadas e maceradas, as amostras foram incubadas à 30o C por cerca de 72 horas, quando se observou crescimento de microrganismos nos meios de cultura: Agar nutriente, meio TSA (Tryptone Soya Agar) e meio de King. A verificação da capacidade de degradação de petróleo e derivados foi realizada em placas de ELISA, expondo a bactéria a uma solução contendo meio de cultura Mínimo, solução do corante DCPIP (sal de sódio de 2,6-dicloroindofenol) e petróleo ou derivado testado (óleo queimado, óleo lubrificante, óleo diesel e gasolina). A leitura positiva para degradação foi observada pela descoloração do DCPIP. Entre nove bactérias testadas, três apresentaram atividade degradativa em diferentes frações de petróleo, óleo diesel e gasolina, e as demais apresentaram variados perfis. Este resultado mostra a potencialidade desses microrganismos para aplicação em processos de biorremediação. A produção de AIA foi testada em meio TSA acrescido com triptofano e somente uma amostra apresentou resultado positivo, enquanto nenhuma foi considerada solubilizadora de fosfatos. Para teste de fixação de nitrogênio os isolados foram inoculados em meios livres de nitrogênio, apresentando crescimento após 24 horas de incubação, no entanto não apresentaram resultados positivos no teste de redução do acetileno, que confirma atividade da enzima nitrogenase. As amostras apresentaram variados perfis para a produção e atuação das bacteriocinas frente aos isolados clínicos testados. Não foi detectada atividade das enzimas endoglucanase, celulolítica e pectinolítica, mas sim de esterases, ß- glucosidases, proteases e amilases. Alguns isolados foram identificados a partir do sequenciamento da região 16S DNA, cuja sequência foi comparada a um banco de dados (GenBank) e então diagnosticados como Bacillus cereus, Staphylococcus pasteuri e Pseudomonas sp. Foi confirmada a caracterização de bactérias endofíticas isoladas de plantas de Cerrado com potencial para utilização em biorremediação, promoção de crescimento vegetal e diversa produção enzimática, com potencialidades para utilização em indústria farmacêutica, alimentícia, têxtil, entre outras.Made available in DSpace on 2014-07-29T15:30:35Z (GMT). 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dc.title.por.fl_str_mv Seleção de microrganismos endofíticos com potencialidades para a biorremediação de ambientes contaminados com hidrocarbonetos de petróleo e/ou derivados
dc.title.alternative.eng.fl_str_mv Selection of endophytic microorganisms for biorremediation in impacted soils with petroleum hydrocarbons and derivates
title Seleção de microrganismos endofíticos com potencialidades para a biorremediação de ambientes contaminados com hidrocarbonetos de petróleo e/ou derivados
spellingShingle Seleção de microrganismos endofíticos com potencialidades para a biorremediação de ambientes contaminados com hidrocarbonetos de petróleo e/ou derivados
OLIVEIRA, Natalia Carvalhaes de
microrganismos endofíticos
biorremediação
enzimas
atividade antimicrobiana
endophytic microorganisms
bioremediation
enzymes
antimicrobial activity
CNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIA
title_short Seleção de microrganismos endofíticos com potencialidades para a biorremediação de ambientes contaminados com hidrocarbonetos de petróleo e/ou derivados
title_full Seleção de microrganismos endofíticos com potencialidades para a biorremediação de ambientes contaminados com hidrocarbonetos de petróleo e/ou derivados
title_fullStr Seleção de microrganismos endofíticos com potencialidades para a biorremediação de ambientes contaminados com hidrocarbonetos de petróleo e/ou derivados
title_full_unstemmed Seleção de microrganismos endofíticos com potencialidades para a biorremediação de ambientes contaminados com hidrocarbonetos de petróleo e/ou derivados
title_sort Seleção de microrganismos endofíticos com potencialidades para a biorremediação de ambientes contaminados com hidrocarbonetos de petróleo e/ou derivados
author OLIVEIRA, Natalia Carvalhaes de
author_facet OLIVEIRA, Natalia Carvalhaes de
author_role author
dc.contributor.advisor1.fl_str_mv VIEIRA, José Daniel Gonçalves
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/1742731776579730
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/7560948180074168
dc.contributor.author.fl_str_mv OLIVEIRA, Natalia Carvalhaes de
contributor_str_mv VIEIRA, José Daniel Gonçalves
dc.subject.por.fl_str_mv microrganismos endofíticos
biorremediação
enzimas
atividade antimicrobiana
topic microrganismos endofíticos
biorremediação
enzimas
atividade antimicrobiana
endophytic microorganisms
bioremediation
enzymes
antimicrobial activity
CNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIA
dc.subject.eng.fl_str_mv endophytic microorganisms
bioremediation
enzymes
antimicrobial activity
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIA
description Endophytic microorganisms live inside plants showing no apparently damage for the host, often assisting in survival of plants, helping its growth with production of phytohormones, phosphates solubilization, nitrogen fixation and enzymes production, or they can metabolize organic contaminants, like petroleum and derivates. This work aimed to isolated and identified endophytic microorganisms of plants present in impacted areas, as well as test their ability in petroleum and its derivatives degradation, identify bacteriocin production, to test their nitrogen fixation capability, phosphate solubilization, indol-acetic acid (IAA) and enzymes production. Plant samples were collected, in an area impacted with asphaltic and mud, were superficially disinfected using 70% ethanol, sodium hypochlorite and sterile distilled water. After macerated and fragmented, the samples were incubated at 30°C for about 72 hours, when growth of microorganisms was observed in culture media: Nutrient Agar, TSA (Tryptone Soya Agar) and King medium. The verification of petroleum and derivatives degradation capacity was performed in ELISA plates, exposing the bacteria to a solution of Minimal Medium, the dye DCPIP solution (2,6-dicloroindofenol sodium salt) and petroleum or derivative tested (burning oil, lubricating oil, diesel oil and gasoline). A positive reading for degradation was observed by discoloration of DCPIP. Among nine bacteria tested, three showed degradative activity in different fractions of petroleum, diesel oil and gasoline, and others showed different profiles. This result shows the potential of these microorganisms for application in bioremediation processes. The production of IAA was tested on TSA medium supplemented with tryptophan and only one sample was positive, while none was considered phosphate solubilizing. Testing the nitrogen fixing, samples were inoculated for a free nitrogen culture media, showing growth after 24 hours of incubation (positive results), however it s isolated did not show positive results in the acetylene reduction test, which confirms activity of the enzyme nitrogenase. The samples showed different profiles for the production and activity of bacteriocins against the clinical isolates tested. There were no detected activity of endoglucanase, cellulolytic and pectinolytic enzymes, but esterases, ß-glucosidases, proteases and amylases were positive. Some isolates were identified from the sequencing of 16S DNA, whose sequence was compared to database (GenBank), and then diagnosed as Bacillus cereus, Staphylococcus pasteuri and Pseudomonas sp. The endophytic bacteria isolated from Cerrado plants confirmed their potential for use in bioremediation, plant growth promotion and diverse enzyme production, with potential for use in pharmaceutical, food, textiles, among others.
publishDate 2009
dc.date.issued.fl_str_mv 2009-03-16
dc.date.available.fl_str_mv 2011-05-02
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dc.identifier.citation.fl_str_mv OLIVEIRA, Natalia Carvalhaes de. Selection of endophytic microorganisms for biorremediation in impacted soils with petroleum hydrocarbons and derivates. 2009. 96 f. Dissertação (Mestrado em Medicina) - Universidade Federal de Goiás, Goiânia, 2009.
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identifier_str_mv OLIVEIRA, Natalia Carvalhaes de. Selection of endophytic microorganisms for biorremediation in impacted soils with petroleum hydrocarbons and derivates. 2009. 96 f. Dissertação (Mestrado em Medicina) - Universidade Federal de Goiás, Goiânia, 2009.
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