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
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/19419
Acceder
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 Técnicas2025-10-15T14:20:33Zoai: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:34982025-10-15 14:20:33.439CONICET 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
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score 13.22299

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