Development of oxidoreductase based electrochemical biosensors

Detalhes bibliográficos
Autor(a) principal: Rodrigues, Patrícia Raquel dos Santos
Data de Publicação: 2013
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/10427
Resumo: Dissertação para obtenção do Grau de Mestre em Biotecnologia
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spelling Development of oxidoreductase based electrochemical biosensorsBiosensorsCytochrome c nitrite reductaseCytochrome P4501A2Sol-gelCarbon nanotubesElectrophoretic depositionDissertação para obtenção do Grau de Mestre em BiotecnologiaThis thesis is divided in 2 sections, each describing the development of an oxidoreductase based biosensor. In the first part human Cytochrome P450 1A2 (CYP1A2) electrochemistry was studied, while the second is focused on the optimization of immobilization platforms and operation methods for amperometric biosensors, using cytochrome c nitrite reductase (ccNiR), (Desulfovibrio desulfuricans ATCC 27774) as a model enzyme. The direct electrochemistry of P450s immobilized in water-based sol-gel thin films was described for the first time. The optimization of the film showed that only the combination of the inorganic matrix and the PEG400 enabled the direct electron transfer reaction and electrocatalytic activity towards oxygen. The amount of dissolved oxygen in solution revealed itself a significant feature in CYP’s electrochemistry – in anaerobic conditions, when small amounts of oxygen are added the PFeIII=II signal’s intensity increased, while in aerobic conditions it disappeared; probably PFeIII is not being regenerated. However, this was not observed with the CYPOR complex, indicating that the reductase has an essential role in the CYP’s catalytic cycle completion; this was also sustained by the fact that only in its presence organic substrates catalysis (caffeine) occurs. The hybrid sol-gel developed for CYP, was optimized for a nitrite biosensor. ccNiR was successfully incorporated while promptly displaying catalytic currents. Although the bioelectrode’s response decreases after day one, it was able to maintain a reasonable catalytic activity over a time span of 2 weeks. Another electrode modification strategy, studied with ccNiR, was based on the electrophoretic deposition of macroporous assemblies of single-walled carbon nanotubes. The macroporous structure was created as a result of the presence of polystyrene beads co-deposited with the carbon nanotubes. An increase in the amount of material was correlated with a higher enzyme activity. Finally, an oxygen scavenger system consisting of glucose oxidase, glucose, and catalase was employed for oxygen removal in an open electrochemical cell. The system completely removed oxygen for over 1 h and was successfully applied to a ccNiR based nitrite sensor.Faculdade de Ciências e TecnologiaAlmeida, Maria GabrielaPereira, SofiaRUNRodrigues, Patrícia Raquel dos Santos2013-09-26T09:09:30Z20132013-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/10427TID:201755602enginfo: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:RCAAP2024-03-11T03:44:12Zoai:run.unl.pt:10362/10427Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:19:28.395721Repositório 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 Development of oxidoreductase based electrochemical biosensors
title Development of oxidoreductase based electrochemical biosensors
spellingShingle Development of oxidoreductase based electrochemical biosensors
Rodrigues, Patrícia Raquel dos Santos
Biosensors
Cytochrome c nitrite reductase
Cytochrome P4501A2
Sol-gel
Carbon nanotubes
Electrophoretic deposition
title_short Development of oxidoreductase based electrochemical biosensors
title_full Development of oxidoreductase based electrochemical biosensors
title_fullStr Development of oxidoreductase based electrochemical biosensors
title_full_unstemmed Development of oxidoreductase based electrochemical biosensors
title_sort Development of oxidoreductase based electrochemical biosensors
author Rodrigues, Patrícia Raquel dos Santos
author_facet Rodrigues, Patrícia Raquel dos Santos
author_role author
dc.contributor.none.fl_str_mv Almeida, Maria Gabriela
Pereira, Sofia
RUN
dc.contributor.author.fl_str_mv Rodrigues, Patrícia Raquel dos Santos
dc.subject.por.fl_str_mv Biosensors
Cytochrome c nitrite reductase
Cytochrome P4501A2
Sol-gel
Carbon nanotubes
Electrophoretic deposition
topic Biosensors
Cytochrome c nitrite reductase
Cytochrome P4501A2
Sol-gel
Carbon nanotubes
Electrophoretic deposition
description Dissertação para obtenção do Grau de Mestre em Biotecnologia
publishDate 2013
dc.date.none.fl_str_mv 2013-09-26T09:09:30Z
2013
2013-01-01T00:00:00Z
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/10362/10427
TID:201755602
url http://hdl.handle.net/10362/10427
identifier_str_mv TID:201755602
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
institution RCAAP
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
repository.mail.fl_str_mv
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