Electrochemical characterization of Dps, a DNA-protecting protein

Detalhes bibliográficos
Autor(a) principal: Pereira, Joana Fernandes Alberto Wilton
Data de Publicação: 2011
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/7820
Resumo: Dissertação para obtenção do Grau de Mestre em Biotecnologia
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spelling Electrochemical characterization of Dps, a DNA-protecting proteinBioelectrochemistryDpsPseudomonas nautica 617Stationary phaseDissertação para obtenção do Grau de Mestre em BiotecnologiaThe objectives of the present work were the purification and electrochemical characterization of Dps and study its iron incorporation mechanism in the presence of hydrogen peroxide. A bacterial miniferritin (Dps) from Pseudomonas (Ps.) nautica 617 was expressed in BL21(DE3) E. coli cells transformed with pET21c(+)-1dps plasmid vector. Protein overexpression was performed in nutritionally rich LB medium containing 100 μg/mL ampicillin. Recombinant Dps was purified through a two-step process: weak ionic exchange chromatography (DEAE Sepharose Fast Flow XK 26/40 resin) and strong ionic exchange chromatography (Q Resource resin). Protein concentration was determined through UV/Visible spectroscopy. The electrochemical study was performed through cyclic voltammetry, square wave voltammetry and chronoamperometry; Dps was either in solution or adsorbed to the working electrode. The chosen supporting electrolyte was 200 mM MOPS pH 7.1 buffer with 200 mM NaCl. Electrochemical assays were performed in aerobic and anaerobic environment and in the presence of Dps and Fe(II) and H2O2, both of Dps co-substrates. For the different assays, working electrodes made of gold, glassy carbon and graphite were used with different results and conclusions. The iron incorporation in the ferroxidase centers and mineral core formation were studied. Assay results were obtained through direct electron transfer. Among the observed results, iron oxidation during protein incubation and oxygen production during the catalytic mechanism were observed. DNA-Dps interaction results were inconclusive, although some unconfirmed evidence exists of DNA protection by Dps from electrochemical oxidation.Faculdade de Ciências e TecnologiaSilva, CristinaTavares, PedroRUNPereira, Joana Fernandes Alberto Wilton2012-09-12T14:53:56Z20112011-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/7820enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T03:40:02Zoai:run.unl.pt:10362/7820Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:17:45.170899Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Electrochemical characterization of Dps, a DNA-protecting protein
title Electrochemical characterization of Dps, a DNA-protecting protein
spellingShingle Electrochemical characterization of Dps, a DNA-protecting protein
Pereira, Joana Fernandes Alberto Wilton
Bioelectrochemistry
Dps
Pseudomonas nautica 617
Stationary phase
title_short Electrochemical characterization of Dps, a DNA-protecting protein
title_full Electrochemical characterization of Dps, a DNA-protecting protein
title_fullStr Electrochemical characterization of Dps, a DNA-protecting protein
title_full_unstemmed Electrochemical characterization of Dps, a DNA-protecting protein
title_sort Electrochemical characterization of Dps, a DNA-protecting protein
author Pereira, Joana Fernandes Alberto Wilton
author_facet Pereira, Joana Fernandes Alberto Wilton
author_role author
dc.contributor.none.fl_str_mv Silva, Cristina
Tavares, Pedro
RUN
dc.contributor.author.fl_str_mv Pereira, Joana Fernandes Alberto Wilton
dc.subject.por.fl_str_mv Bioelectrochemistry
Dps
Pseudomonas nautica 617
Stationary phase
topic Bioelectrochemistry
Dps
Pseudomonas nautica 617
Stationary phase
description Dissertação para obtenção do Grau de Mestre em Biotecnologia
publishDate 2011
dc.date.none.fl_str_mv 2011
2011-01-01T00:00:00Z
2012-09-12T14:53:56Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/7820
url http://hdl.handle.net/10362/7820
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Faculdade de Ciências e Tecnologia
publisher.none.fl_str_mv Faculdade de Ciências e Tecnologia
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
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reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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