Caracteriza????o do potencial de degrada????o de um isolado bacteriano oriundo da regi??o Amaz??nica

• Author(s): Castro, Diogo Pereira de
• Date: 2018
• Origin: Oasisbr
• Subject(s): Petr??leo; Biorremedia????o; Degrada????o de Hidrocarbonetos; Prote??mica; CIENCIAS BIOLOGICAS
• Document Type: Doctoral thesis
• Language: Portuguese

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FAPEAM - Funda????o de Amparo ?? Pesquisa do Estado do Amazonas

Bioremediation is the use of microorganisms capable of metabolizing contaminants and converts them into less toxic products. In previous studies, we performed bacteria isolation from contaminated area for oil waste in Amazon. Despite its oil degradation potential, the application of bacteria in contaminated sites may interfere with their ecological balance. This study aimed to promote studies of gene characterization and hydrocarbon degradation potential with a selected bacterial strain and identify and define the proteins involved in degradation process. To achieve this objectives degraded substances have been identified by gas chromatography and proteins involved by proteomics; Genome analysis allowed characterization of genes related to xenobiotics degradation. The strain was identified as Pseudomonas putida S16. 51 substances were detected by gas chromatography, of these, 23 were completely degraded, including PAH containing naphthalene (100%), anthracene (11.49%) and phenanthrene (7.41%). We identified 89 genes related to xenobiotic degradation in P. putida S16 AM genome, genes capable of degrading naphthalene, anthracene and phenanthrene (nah, phn, fdx, catA), cytochrome production and chemotaxis. In proteome analysis, proteins were expressed oxidation-reduction, metabolic processes, polyamines, aldehyde dehydrogenase, carbon storage, chemotaxis and amino acid synthesis. 355 proteins were identified with redundancy expressed during 7h contact to P. putida S16 AM (I4) with petroleum, and 268 during 75h contact; For glycerol, 467 were identified in 7h interaction, and 228 in 75h contact. The presence of proteins related to metabolism and oxidationreduction when the strain under study was in contact to the petroleum suggest the use of this as a carbon source for bacterial metabolism. These studies are important because of identification of genes and proteins with potential to become practical application of biotechnology products.

A biorremedia????o ?? a utiliza????o de microrganismos capazes de metabolizar contaminantes e transforma-los em produtos menos t??xicos. Em estudos anteriores, foi realizado o isolamento de bact??rias provenientes de ??rea contaminada por res??duos de petr??leo na Amaz??nia. Apesar de possu??rem o potencial de degrada????o de petr??leo, a aplica????o de bact??rias em ambientes contaminados pode interferir em seu equil??brio ecol??gico. Este estudo prop??s-se a sele????o de uma linhagem bacteriana para promover estudos de caracteriza????o g??nica, identifica????o do potencial de degrada????o de hidrocarbonetos e defini????o das prote??nas envolvidas no processo de degrada????o. Para alcan??ar os objetivos, foram identificadas subst??ncias degradadas por cromatografia gasosa e prote??nas envolvidas por prote??mica; A an??lise do genoma permitiu a caracteriza????o dos genes relacionados a degrada????o de compostos xenobi??ticos atrav??s da revis??o da literatura. A cepa foi identificada como Pseudomonas putida S16. Foram detectadas 51 subst??ncias por cromatografia gasosa, destas, 23 foram completamente degradadas, incluindo HPAs contendo naftaleno (100%), antraceno (11,49%) e fenantreno (7,41%). Foram identificados 89 genes relacionados a degrada????o de xenob??oticos no genoma da P. putida S16 AM, genes capazes de degradar naftaleno, antraceno e fenantreno (nah, phn, fdx, catA), produ????o de citocromo e quimiotaxia. Na an??lise prote??mica, foram expressas prote??nas de ??xido-redu????o, processos metab??licos, poliaminas, alde??do desidrogenase, armazenamento de carbono, quimiotaxia e s??ntese de amino??cidos. Foram identificadas 355 prote??nas com redund??ncia expressas durante o contato de 7h da P. putida S16 AM (I4) com o petr??leo, e 268 no contato de 75h; Para o glicerol, foram identificadas 467 na intera????o de 7h, e 228 no contato de 75h. A presen??a de prote??nas relacionadas ao metabolismo e ??xidoredu????o quando a cepa em estudo esteve em contato com o petr??leo evidenciam a utiliza????o deste como fonte de carbono para o metabolismo bacteriano. Estes estudos s??o importantes devido ?? identifica????o de genes e prote??nas que possuem o potencial de se tornarem produtos biotecnol??gicos de aplica????o pr??tica.