Purification and Characterization of an Alkaline Phosphatase Induced by Phosphorus Starvation in Common Bean (Phaseolus vulgaris L.) Roots

Two phosphatase isoforms from roots of the common bean(Phaseolus vulgaris L.) showed an increase in activity in responseto phosphate deficiency. One of them (APIII) was chosen for further purification through ionic exchange chromatography and preparative electrophoresis. The estimated molecular mass...

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Detalles Bibliográficos
Autores: Lorena Morales, Natalia Gutiérrez, Vanessa Maya, Carmen Parra, Eleazar Martínez-Barajas, Patricia Coello
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2012
País:México
Institución:Universidad Nacional Autónoma de México
Repositorio:Redalyc-UNAM
OAI Identifier:oai:redalyc.org:47523284013
Acceso en línea:https://www.redalyc.org/articulo.oa?id=47523284013
Access Level:acceso abierto
Palabra clave:Química
purification
pyrophosphate
Phaseolus vulgaris
Alkaline phosphatase
phosphate deficiency
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spelling Purification and Characterization of an Alkaline Phosphatase Induced by Phosphorus Starvation in Common Bean (Phaseolus vulgaris L.) RootsLorena MoralesNatalia GutiérrezVanessa MayaCarmen ParraEleazar Martínez-BarajasPatricia CoelloQuímicapurificationpyrophosphatePhaseolus vulgarisAlkaline phosphatasephosphate deficiencyTwo phosphatase isoforms from roots of the common bean(Phaseolus vulgaris L.) showed an increase in activity in responseto phosphate deficiency. One of them (APIII) was chosen for further purification through ionic exchange chromatography and preparative electrophoresis. The estimated molecular mass of APIII was 35 kDa by both SDS-PAGE and gel filtration analyses, suggesting a monomeric form of the active enzyme. The phosphatase was classified as an alkaline phosphatase based on the requirement of pH 8 for optimum catalysis. It not only exhibited broad substrate specificity, with the most activity against pyrophosphate, but also effectively catalyzed the hydrolysis of polyphosphate, glucose-1-phosphate and phosphoenolpyruvate. Activity was completely inhibited by molybdate, vanadate and phosphate but was only partially inhibited by fluoride. Although divalent cations were not essential for the pyrophosphatase activity of this enzyme, the hydrolysis of pyrophosphate increased substantially in the presence of Mg2+.Sociedad Química de México2012info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdf1870-249Xhttps://www.redalyc.org/articulo.oa?id=47523284013Journal of the Mexican Chemical Society (México) Num.1 Vol.56reponame:Redalyc-UNAMinstname:Universidad Nacional Autónoma de Méxicoinstacron:UNAMenhttp://www.redalyc.org/revista.oa?id=475Journal of the Mexican Chemical Societyinfo:eu-repo/semantics/openAccessoai:redalyc.org:475232840132025-09-03T18:05:45Z
dc.title.none.fl_str_mv Purification and Characterization of an Alkaline Phosphatase Induced by Phosphorus Starvation in Common Bean (Phaseolus vulgaris L.) Roots
title Purification and Characterization of an Alkaline Phosphatase Induced by Phosphorus Starvation in Common Bean (Phaseolus vulgaris L.) Roots
spellingShingle Purification and Characterization of an Alkaline Phosphatase Induced by Phosphorus Starvation in Common Bean (Phaseolus vulgaris L.) Roots
Lorena Morales
Química
purification
pyrophosphate
Phaseolus vulgaris
Alkaline phosphatase
phosphate deficiency
title_short Purification and Characterization of an Alkaline Phosphatase Induced by Phosphorus Starvation in Common Bean (Phaseolus vulgaris L.) Roots
title_full Purification and Characterization of an Alkaline Phosphatase Induced by Phosphorus Starvation in Common Bean (Phaseolus vulgaris L.) Roots
title_fullStr Purification and Characterization of an Alkaline Phosphatase Induced by Phosphorus Starvation in Common Bean (Phaseolus vulgaris L.) Roots
title_full_unstemmed Purification and Characterization of an Alkaline Phosphatase Induced by Phosphorus Starvation in Common Bean (Phaseolus vulgaris L.) Roots
title_sort Purification and Characterization of an Alkaline Phosphatase Induced by Phosphorus Starvation in Common Bean (Phaseolus vulgaris L.) Roots
dc.creator.none.fl_str_mv Lorena Morales
Natalia Gutiérrez
Vanessa Maya
Carmen Parra
Eleazar Martínez-Barajas
Patricia Coello
author Lorena Morales
author_facet Lorena Morales
Natalia Gutiérrez
Vanessa Maya
Carmen Parra
Eleazar Martínez-Barajas
Patricia Coello
author_role author
author2 Natalia Gutiérrez
Vanessa Maya
Carmen Parra
Eleazar Martínez-Barajas
Patricia Coello
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Química
purification
pyrophosphate
Phaseolus vulgaris
Alkaline phosphatase
phosphate deficiency
topic Química
purification
pyrophosphate
Phaseolus vulgaris
Alkaline phosphatase
phosphate deficiency
description Two phosphatase isoforms from roots of the common bean(Phaseolus vulgaris L.) showed an increase in activity in responseto phosphate deficiency. One of them (APIII) was chosen for further purification through ionic exchange chromatography and preparative electrophoresis. The estimated molecular mass of APIII was 35 kDa by both SDS-PAGE and gel filtration analyses, suggesting a monomeric form of the active enzyme. The phosphatase was classified as an alkaline phosphatase based on the requirement of pH 8 for optimum catalysis. It not only exhibited broad substrate specificity, with the most activity against pyrophosphate, but also effectively catalyzed the hydrolysis of polyphosphate, glucose-1-phosphate and phosphoenolpyruvate. Activity was completely inhibited by molybdate, vanadate and phosphate but was only partially inhibited by fluoride. Although divalent cations were not essential for the pyrophosphatase activity of this enzyme, the hydrolysis of pyrophosphate increased substantially in the presence of Mg2+.
publishDate 2012
dc.date.none.fl_str_mv 2012
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv 1870-249X
https://www.redalyc.org/articulo.oa?id=47523284013
identifier_str_mv 1870-249X
url https://www.redalyc.org/articulo.oa?id=47523284013
dc.language.none.fl_str_mv en
language_invalid_str_mv en
dc.relation.none.fl_str_mv http://www.redalyc.org/revista.oa?id=475
dc.rights.none.fl_str_mv Journal of the Mexican Chemical Society
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Journal of the Mexican Chemical Society
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Sociedad Química de México
publisher.none.fl_str_mv Sociedad Química de México
dc.source.none.fl_str_mv Journal of the Mexican Chemical Society (México) Num.1 Vol.56
reponame:Redalyc-UNAM
instname:Universidad Nacional Autónoma de México
instacron:UNAM
instname_str Universidad Nacional Autónoma de México
instacron_str UNAM
institution UNAM
reponame_str Redalyc-UNAM
collection Redalyc-UNAM
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 14.965132

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