Development of oxidoreductase based electrochemical biosensors
Autor(a) principal: | |
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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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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 |
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1799137837926318080 |