The homopentameric chlorite dismutase from Magnetospirillum sp
- Autores
- Freire, Diana M.; Rivas, Maria Gabriela; Dias, André M.; Lopes, Ana T.; Costa, Cristina; Santos Silva, Teresa; Van Doorslaer, Sabine; González, Pablo Javier
- Año de publicación
- 2015
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- Chlorite dismutase (Cld) is a b-type hemecontaining enzymethat catalyzes the reduction of chlorite into chloride plus dioxygen. This enzyme has gained attention because it can be used in the development of bioremediation processes, biosensors, and controlled dioxygen production. In the present work, Cld was purified from Magnetospirillum sp. cells cultured anaerobically with acetate/perchlorate until stationary phase. Biochemical, spectroscopic and X-ray crystallography methods showed that Cld from Magnetospirillum sp. is a ~140 kDa homopentamer comprising ~27.8 kDa monomers. Preliminary X-ray crystallography studies confirmed the quaternary structure and the presence of one b-type heme per monomer. The EPR spectroscopic signature of the as-purified Cld samples is affected by the buffer composition used during the purification. Potassium phosphate buffer is the only buffer that affected neither the spectral nor the kinetic properties of Cld. Kinetic studies in solution revealed that Cld from Magnetospirillum sp. decomposes chlorite at high turnover rates with optimal pH 6.0. A temperature below 10 °C is required to avoid enzyme inactivation due to cofactor bleaching during turnover, and to achieve full substrate consumption. Cld kinetic parameters were not affected when kinetic assays were performed in the presence of air or under argon atmosphere, but chloride is a weak mixed inhibitor that modifies the EPR signal of as-prepared samples.
Fil: Freire, Diana M.. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal
Fil: Rivas, Maria Gabriela. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina
Fil: Dias, André M.. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal
Fil: Lopes, Ana T.. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal
Fil: Costa, Cristina. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal
Fil: Santos Silva, Teresa. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal
Fil: Van Doorslaer, Sabine. University of Antwerp. Department of Physics, ; Bélgica
Fil: González, Pablo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal - Materia
-
Chlorite Dismutase
Enzyme Kinetics
X-Ray Crystallography
Epr Spectroscopy
Magnetospirillum
Perchlorate-Reducing Bacteria - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/19419
Ver los metadatos del registro completo
id |
CONICETDig_7391cfb612a75a3453839b88be58aa21 |
---|---|
oai_identifier_str |
oai:ri.conicet.gov.ar:11336/19419 |
network_acronym_str |
CONICETDig |
repository_id_str |
3498 |
network_name_str |
CONICET Digital (CONICET) |
spelling |
The homopentameric chlorite dismutase from Magnetospirillum spFreire, Diana M.Rivas, Maria GabrielaDias, André M.Lopes, Ana T.Costa, CristinaSantos Silva, TeresaVan Doorslaer, SabineGonzález, Pablo JavierChlorite DismutaseEnzyme KineticsX-Ray CrystallographyEpr SpectroscopyMagnetospirillumPerchlorate-Reducing Bacteriahttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Chlorite dismutase (Cld) is a b-type hemecontaining enzymethat catalyzes the reduction of chlorite into chloride plus dioxygen. This enzyme has gained attention because it can be used in the development of bioremediation processes, biosensors, and controlled dioxygen production. In the present work, Cld was purified from Magnetospirillum sp. cells cultured anaerobically with acetate/perchlorate until stationary phase. Biochemical, spectroscopic and X-ray crystallography methods showed that Cld from Magnetospirillum sp. is a ~140 kDa homopentamer comprising ~27.8 kDa monomers. Preliminary X-ray crystallography studies confirmed the quaternary structure and the presence of one b-type heme per monomer. The EPR spectroscopic signature of the as-purified Cld samples is affected by the buffer composition used during the purification. Potassium phosphate buffer is the only buffer that affected neither the spectral nor the kinetic properties of Cld. Kinetic studies in solution revealed that Cld from Magnetospirillum sp. decomposes chlorite at high turnover rates with optimal pH 6.0. A temperature below 10 °C is required to avoid enzyme inactivation due to cofactor bleaching during turnover, and to achieve full substrate consumption. Cld kinetic parameters were not affected when kinetic assays were performed in the presence of air or under argon atmosphere, but chloride is a weak mixed inhibitor that modifies the EPR signal of as-prepared samples.Fil: Freire, Diana M.. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; PortugalFil: Rivas, Maria Gabriela. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Dias, André M.. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; PortugalFil: Lopes, Ana T.. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; PortugalFil: Costa, Cristina. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; PortugalFil: Santos Silva, Teresa. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; PortugalFil: Van Doorslaer, Sabine. University of Antwerp. Department of Physics, ; BélgicaFil: González, Pablo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; PortugalElsevier2015-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/19419Freire, Diana M.; Rivas, Maria Gabriela; Dias, André M.; Lopes, Ana T.; Costa, Cristina; et al.; The homopentameric chlorite dismutase from Magnetospirillum sp; Elsevier; Journal of Inorganic Biochemistry; 151; 7-2015; 1-90162-0134CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0162013415300295info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jinorgbio.2015.07.006info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:32:37Zoai:ri.conicet.gov.ar:11336/19419instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:32:37.624CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
The homopentameric chlorite dismutase from Magnetospirillum sp |
title |
The homopentameric chlorite dismutase from Magnetospirillum sp |
spellingShingle |
The homopentameric chlorite dismutase from Magnetospirillum sp Freire, Diana M. Chlorite Dismutase Enzyme Kinetics X-Ray Crystallography Epr Spectroscopy Magnetospirillum Perchlorate-Reducing Bacteria |
title_short |
The homopentameric chlorite dismutase from Magnetospirillum sp |
title_full |
The homopentameric chlorite dismutase from Magnetospirillum sp |
title_fullStr |
The homopentameric chlorite dismutase from Magnetospirillum sp |
title_full_unstemmed |
The homopentameric chlorite dismutase from Magnetospirillum sp |
title_sort |
The homopentameric chlorite dismutase from Magnetospirillum sp |
dc.creator.none.fl_str_mv |
Freire, Diana M. Rivas, Maria Gabriela Dias, André M. Lopes, Ana T. Costa, Cristina Santos Silva, Teresa Van Doorslaer, Sabine González, Pablo Javier |
author |
Freire, Diana M. |
author_facet |
Freire, Diana M. Rivas, Maria Gabriela Dias, André M. Lopes, Ana T. Costa, Cristina Santos Silva, Teresa Van Doorslaer, Sabine González, Pablo Javier |
author_role |
author |
author2 |
Rivas, Maria Gabriela Dias, André M. Lopes, Ana T. Costa, Cristina Santos Silva, Teresa Van Doorslaer, Sabine González, Pablo Javier |
author2_role |
author author author author author author author |
dc.subject.none.fl_str_mv |
Chlorite Dismutase Enzyme Kinetics X-Ray Crystallography Epr Spectroscopy Magnetospirillum Perchlorate-Reducing Bacteria |
topic |
Chlorite Dismutase Enzyme Kinetics X-Ray Crystallography Epr Spectroscopy Magnetospirillum Perchlorate-Reducing Bacteria |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
dc.description.none.fl_txt_mv |
Chlorite dismutase (Cld) is a b-type hemecontaining enzymethat catalyzes the reduction of chlorite into chloride plus dioxygen. This enzyme has gained attention because it can be used in the development of bioremediation processes, biosensors, and controlled dioxygen production. In the present work, Cld was purified from Magnetospirillum sp. cells cultured anaerobically with acetate/perchlorate until stationary phase. Biochemical, spectroscopic and X-ray crystallography methods showed that Cld from Magnetospirillum sp. is a ~140 kDa homopentamer comprising ~27.8 kDa monomers. Preliminary X-ray crystallography studies confirmed the quaternary structure and the presence of one b-type heme per monomer. The EPR spectroscopic signature of the as-purified Cld samples is affected by the buffer composition used during the purification. Potassium phosphate buffer is the only buffer that affected neither the spectral nor the kinetic properties of Cld. Kinetic studies in solution revealed that Cld from Magnetospirillum sp. decomposes chlorite at high turnover rates with optimal pH 6.0. A temperature below 10 °C is required to avoid enzyme inactivation due to cofactor bleaching during turnover, and to achieve full substrate consumption. Cld kinetic parameters were not affected when kinetic assays were performed in the presence of air or under argon atmosphere, but chloride is a weak mixed inhibitor that modifies the EPR signal of as-prepared samples. Fil: Freire, Diana M.. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal Fil: Rivas, Maria Gabriela. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina Fil: Dias, André M.. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal Fil: Lopes, Ana T.. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal Fil: Costa, Cristina. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal Fil: Santos Silva, Teresa. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal Fil: Van Doorslaer, Sabine. University of Antwerp. Department of Physics, ; Bélgica Fil: González, Pablo Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidade Nova de Lisboa. Faculdade de Ciências e Tecnologia. Departamento de Química; Portugal |
description |
Chlorite dismutase (Cld) is a b-type hemecontaining enzymethat catalyzes the reduction of chlorite into chloride plus dioxygen. This enzyme has gained attention because it can be used in the development of bioremediation processes, biosensors, and controlled dioxygen production. In the present work, Cld was purified from Magnetospirillum sp. cells cultured anaerobically with acetate/perchlorate until stationary phase. Biochemical, spectroscopic and X-ray crystallography methods showed that Cld from Magnetospirillum sp. is a ~140 kDa homopentamer comprising ~27.8 kDa monomers. Preliminary X-ray crystallography studies confirmed the quaternary structure and the presence of one b-type heme per monomer. The EPR spectroscopic signature of the as-purified Cld samples is affected by the buffer composition used during the purification. Potassium phosphate buffer is the only buffer that affected neither the spectral nor the kinetic properties of Cld. Kinetic studies in solution revealed that Cld from Magnetospirillum sp. decomposes chlorite at high turnover rates with optimal pH 6.0. A temperature below 10 °C is required to avoid enzyme inactivation due to cofactor bleaching during turnover, and to achieve full substrate consumption. Cld kinetic parameters were not affected when kinetic assays were performed in the presence of air or under argon atmosphere, but chloride is a weak mixed inhibitor that modifies the EPR signal of as-prepared samples. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-07 |
dc.type.none.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
format |
article |
status_str |
publishedVersion |
dc.identifier.none.fl_str_mv |
http://hdl.handle.net/11336/19419 Freire, Diana M.; Rivas, Maria Gabriela; Dias, André M.; Lopes, Ana T.; Costa, Cristina; et al.; The homopentameric chlorite dismutase from Magnetospirillum sp; Elsevier; Journal of Inorganic Biochemistry; 151; 7-2015; 1-9 0162-0134 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/19419 |
identifier_str_mv |
Freire, Diana M.; Rivas, Maria Gabriela; Dias, André M.; Lopes, Ana T.; Costa, Cristina; et al.; The homopentameric chlorite dismutase from Magnetospirillum sp; Elsevier; Journal of Inorganic Biochemistry; 151; 7-2015; 1-9 0162-0134 CONICET Digital CONICET |
dc.language.none.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0162013415300295 info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jinorgbio.2015.07.006 |
dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-nd/2.5/ar/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/ |
dc.format.none.fl_str_mv |
application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Elsevier |
publisher.none.fl_str_mv |
Elsevier |
dc.source.none.fl_str_mv |
reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
reponame_str |
CONICET Digital (CONICET) |
collection |
CONICET Digital (CONICET) |
instname_str |
Consejo Nacional de Investigaciones Científicas y Técnicas |
repository.name.fl_str_mv |
CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
repository.mail.fl_str_mv |
dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
_version_ |
1844612996596236288 |
score |
13.070432 |