Two superoxide dismutases from TnOtchr are involved in detoxification of reactive oxygen species induced by chromate
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10316/108501 https://doi.org/10.1186/s12866-016-0648-0 |
Resumo: | Background: Superoxide dismutases (SOD) have been reported as the most relevant bacterial enzymes involved in cells protection from reactive oxygen species (ROS). These toxic species are often the product of heavy metal stress. Results: Two genes, chrC and chrF, from TnOtchr genetic determinant of strain Ochrobactrum tritici 5bvl1 were cloned in Escherichia coli in order to overexpress the respective proteins. Both proteins were purified and characterized as superoxide dismutases. ChrC was confirmed as being a Fe-SOD, and the enzymatic activity of the ChrF, not inhibited by hydrogen peroxide or potassium cyanide, suggested its inclusion in the Mn-SOD family. This identification was supported by chemical quantification of total metal content in purified enzyme. Both enzymes showed a maximum activity between pH 7.2-7.5. ChrF retained nearly full activity over a broader range of pH and was slightly more thermostable than ChrC. The genes encoding these enzymes in strain O. tritici 5bvl1 were inactivated, developing single and double mutants, to understand the contribution of these enzymes in detoxification mechanism of reactive oxygen species induced by chromate. During chromate stress, assays using fluorescent dyes indicated an increase of these toxic compounds in chrC, chrF and chrC/chrF mutant cells. Conclusions: In spite of the multiple genes coding for putative superoxide dismutase enzymes detected in the genome of O. tritici 5bvl1, the ChrC and ChrF might help the strain to decrease the levels of reactive oxygen species in cells. |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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7160 |
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Two superoxide dismutases from TnOtchr are involved in detoxification of reactive oxygen species induced by chromateSuperoxide dismutasesReactive oxygen speciesChromate stressMutant cellsFluorescent dyesBacterial ProteinsChromatesOchrobactrumReactive Oxygen SpeciesSuperoxide DismutaseBackground: Superoxide dismutases (SOD) have been reported as the most relevant bacterial enzymes involved in cells protection from reactive oxygen species (ROS). These toxic species are often the product of heavy metal stress. Results: Two genes, chrC and chrF, from TnOtchr genetic determinant of strain Ochrobactrum tritici 5bvl1 were cloned in Escherichia coli in order to overexpress the respective proteins. Both proteins were purified and characterized as superoxide dismutases. ChrC was confirmed as being a Fe-SOD, and the enzymatic activity of the ChrF, not inhibited by hydrogen peroxide or potassium cyanide, suggested its inclusion in the Mn-SOD family. This identification was supported by chemical quantification of total metal content in purified enzyme. Both enzymes showed a maximum activity between pH 7.2-7.5. ChrF retained nearly full activity over a broader range of pH and was slightly more thermostable than ChrC. The genes encoding these enzymes in strain O. tritici 5bvl1 were inactivated, developing single and double mutants, to understand the contribution of these enzymes in detoxification mechanism of reactive oxygen species induced by chromate. During chromate stress, assays using fluorescent dyes indicated an increase of these toxic compounds in chrC, chrF and chrC/chrF mutant cells. Conclusions: In spite of the multiple genes coding for putative superoxide dismutase enzymes detected in the genome of O. tritici 5bvl1, the ChrC and ChrF might help the strain to decrease the levels of reactive oxygen species in cells.Springer Nature2016-03-05info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10316/108501http://hdl.handle.net/10316/108501https://doi.org/10.1186/s12866-016-0648-0eng1471-2180Branco, RitaMorais, Paula V.info: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:RCAAP2023-08-30T11:24:22Zoai:estudogeral.uc.pt:10316/108501Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T21:24:47.512086Repositório 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 |
Two superoxide dismutases from TnOtchr are involved in detoxification of reactive oxygen species induced by chromate |
title |
Two superoxide dismutases from TnOtchr are involved in detoxification of reactive oxygen species induced by chromate |
spellingShingle |
Two superoxide dismutases from TnOtchr are involved in detoxification of reactive oxygen species induced by chromate Branco, Rita Superoxide dismutases Reactive oxygen species Chromate stress Mutant cells Fluorescent dyes Bacterial Proteins Chromates Ochrobactrum Reactive Oxygen Species Superoxide Dismutase |
title_short |
Two superoxide dismutases from TnOtchr are involved in detoxification of reactive oxygen species induced by chromate |
title_full |
Two superoxide dismutases from TnOtchr are involved in detoxification of reactive oxygen species induced by chromate |
title_fullStr |
Two superoxide dismutases from TnOtchr are involved in detoxification of reactive oxygen species induced by chromate |
title_full_unstemmed |
Two superoxide dismutases from TnOtchr are involved in detoxification of reactive oxygen species induced by chromate |
title_sort |
Two superoxide dismutases from TnOtchr are involved in detoxification of reactive oxygen species induced by chromate |
author |
Branco, Rita |
author_facet |
Branco, Rita Morais, Paula V. |
author_role |
author |
author2 |
Morais, Paula V. |
author2_role |
author |
dc.contributor.author.fl_str_mv |
Branco, Rita Morais, Paula V. |
dc.subject.por.fl_str_mv |
Superoxide dismutases Reactive oxygen species Chromate stress Mutant cells Fluorescent dyes Bacterial Proteins Chromates Ochrobactrum Reactive Oxygen Species Superoxide Dismutase |
topic |
Superoxide dismutases Reactive oxygen species Chromate stress Mutant cells Fluorescent dyes Bacterial Proteins Chromates Ochrobactrum Reactive Oxygen Species Superoxide Dismutase |
description |
Background: Superoxide dismutases (SOD) have been reported as the most relevant bacterial enzymes involved in cells protection from reactive oxygen species (ROS). These toxic species are often the product of heavy metal stress. Results: Two genes, chrC and chrF, from TnOtchr genetic determinant of strain Ochrobactrum tritici 5bvl1 were cloned in Escherichia coli in order to overexpress the respective proteins. Both proteins were purified and characterized as superoxide dismutases. ChrC was confirmed as being a Fe-SOD, and the enzymatic activity of the ChrF, not inhibited by hydrogen peroxide or potassium cyanide, suggested its inclusion in the Mn-SOD family. This identification was supported by chemical quantification of total metal content in purified enzyme. Both enzymes showed a maximum activity between pH 7.2-7.5. ChrF retained nearly full activity over a broader range of pH and was slightly more thermostable than ChrC. The genes encoding these enzymes in strain O. tritici 5bvl1 were inactivated, developing single and double mutants, to understand the contribution of these enzymes in detoxification mechanism of reactive oxygen species induced by chromate. During chromate stress, assays using fluorescent dyes indicated an increase of these toxic compounds in chrC, chrF and chrC/chrF mutant cells. Conclusions: In spite of the multiple genes coding for putative superoxide dismutase enzymes detected in the genome of O. tritici 5bvl1, the ChrC and ChrF might help the strain to decrease the levels of reactive oxygen species in cells. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-03-05 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10316/108501 http://hdl.handle.net/10316/108501 https://doi.org/10.1186/s12866-016-0648-0 |
url |
http://hdl.handle.net/10316/108501 https://doi.org/10.1186/s12866-016-0648-0 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
1471-2180 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Springer Nature |
publisher.none.fl_str_mv |
Springer Nature |
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 |
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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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1799134131467059200 |