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Master's Dissertation
DOI
10.11606/D.87.2012.tde-05022013-080751
Document
Author
Full name
Louise Hase Gracioso
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Perpetuo, Elen Aquino (President)
Sales, Claudiana Paula de Souza
Vicente, Elisabete Jose
Title in Portuguese
Análise das proteínas expressas em resposta ao fenol em bactérias isoladas da zona industrial de Cubatão - SP
Keywords in Portuguese
Biodegradação
Compostos fenólicos
Indústrias
Metabolismo celular
Poluição ambiental
Abstract in Portuguese
Os compostos fenólicos pertencem a um grupo tóxico de poluentes ambientais descartados do processo de muitas indústrias tais como refinarias de óleo e indústrias químicas. Embora o fenol possua ação bactericida, alguns micro-organismos adquiriram a habilidade de se adaptar e utilizar este composto como fonte de carbono e energia, através do controle coordenado de vias metabólicas (catabólicas). A expressão destas vias pode ser regulada por: mecanismos de controle globais ou por uma via específica de resposta controlada, porém estes mecanismos ainda não são bem compreendidos. O presente trabalho pretendeu isolar e identificar micro-organismos de um ambiente contaminado para tratamento biológico de efluentes fenólicos, bem como analisar o padrão de proteínas citosólicas expressas em função da exposição à duas diferentes fontes de carbono (glicose ou fenol). As linhagens isoladas de Cubatão-SP foram identificadas pela amplificação e sequenciamento do gene 16S DNAr, resultando em 100 % de similaridade com os gêneros Achromobacter e Pandoraea. Os ensaios de biodegradação em diferentes concentrações de fenol (200 a 600 mg.L-1) mostraram que as duas linhagens foram capazes de degradar 100 % do fenol. As proteínas expressas por Achromobacter sp. em resposta ao fenol foram submetidas a eletroforese 2D e os resultados sugerem que a biodegradação do fenol foi realizada através da meta clivagem do anel aromático, pois três enzimas desta via foram identificadas (proteína degradação do fenol via meta- clivagem, 2- hidroximucônico semialdeído desidrogenase 1 e 4-hidroxi-2-oxovalerate aldolase). Outras enzimas envolvidas no metabolismo celular também foram identificadas, reforçando a hipótese que o fenol altera todo o metabolismo celular, envolvendo as mais diferentes vias metabólicas para que a célula possa superar o estresse celular ocasionado por esta exposição.
Title in English
Analysis of expressed proteins in response to phenol in bacteria isolated from the industrial area of Cubatao-SP.
Keywords in English
Biodegradation
Cell metabolism
Environmental pollution
Industries
Phenolic compounds
Abstract in English
Phenolic compounds belong to a group of toxic environmental pollutants discharged from the process in many industries such as oil refineries and chemical plants. Although phenol has bactericidal action, some microorganisms acquired the ability to adapt and use this compound as a source of carbon and energy, through the coordinated control of metabolic pathways (catabolic). The expression of these pathways may be regulated by: control mechanisms via a global or specific response, but these mechanisms are not well understood. This work aims to isolate and identify micro-organisms from a contaminated environment for biological treatment of phenolic wastewaters, as well as analyzing the pattern of cytosolic proteins expressed in terms of exposure to two different carbon sources (glucose or phenol). The strains isolated from Cubatao-SP were identified by amplification and sequencing of 16S rDNA, resulting in 100% similarity with the genera Achromobacter and Pandoraea. The biodegradation assays at different concentrations of phenol (200 to 600 mg.L-1) showed that both strains were able of degrading 100% of the phenol. The proteins expressed by Achromobacter sp. in response to phenol were subjected to 2D electrophoresis and the results suggest that the biodegradation of phenol was performed using the meta cleavage of the aromatic ring, once three enzymes of this pathway have been identified (protein degradation meta-cleavage pathway phenol, 2-hydroxymuconic semialdehyde dehydrogenase 1 and 4-hydroxy-2-oxovalerate aldolase). Other enzymes involved in cellular metabolism were also identified; reinforcing the hypothesis that phenol modifies the entire cellular metabolism, involving very different metabolic pathways for the cell can overcome stress caused by this exposure.
 
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Publishing Date
2013-02-22
 
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