Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties

Autores
Minchiotti, Mariana Cecilia; Scalambro, Maria Belen; Vargas, Laura Inés; Coronel, Carlos Enrique; Madoery, Ricardo Roman
Año de publicación
2008
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
A protein with phospholipase A2 (PLA2) activity was isolated from soybean (Glycine max) seeds. The affinity column chromatography techniques by using Cibacron Blue and immobilized substrate onto Eupergit C, were important steps in the purification protocol. The electrophoretic mobility showed by the purified enzyme, agreed with a molecular mass of 14 kDa. The PLA2 activity against liposomes was determined by measurement of apparent absorbance changes at 340 nm. Also, high performance thin layer chromatography (HPTLC) analysis confirmed selective hydrolysis at the sn-2 position of soybean phospholipids. PLA2 exhibited millimolar calcium dependence and had slightly alkaline pH optimum. The enzyme was stimulated by auxins and completely inactivated by ammonium sulfate. Furthermore, it was irreversibly inactivated by p-bromo-phenacyl bromide, the specific inhibitor of secretory PLA2s. In addition, Glycine max PLA2 showed high stability against heat treatment and organic solvents. The enzyme showed activity toward multilamellar vesicles of soybean phospholipids, with a Vmax = 950 U/mg and a KM = 0.78 mM. The hyperbolic behavior observed was coherent with a hopping mode of action, one of the two characteristic interfacial mechanisms of PLA2s. All these data agree with the expected properties for a secretory PLA2 being the first soybean enzyme of this type. The new PLA2 developed lipolytic activity on a water in oil microemulsion reaction system, which is suitable for lysophospholipid production. These lysoderivatives are valuable biosurfactants for food and pharmaceutical industries.
Fil: Minchiotti, Mariana Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; Argentina
Fil: Scalambro, Maria Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba. Facultad de Cs.agropecuarias. Departamento de Fundamentación Biologica; Argentina
Fil: Vargas, Laura Inés. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; Argentina
Fil: Coronel, Carlos Enrique. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
Fil: Madoery, Ricardo Roman. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; Argentina
Materia
Phospholipase A2
Properties
Purification
Soybean
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/58410
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic propertiesMinchiotti, Mariana CeciliaScalambro, Maria BelenVargas, Laura InésCoronel, Carlos EnriqueMadoery, Ricardo RomanPhospholipase A2PropertiesPurificationSoybeanhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1A protein with phospholipase A2 (PLA2) activity was isolated from soybean (Glycine max) seeds. The affinity column chromatography techniques by using Cibacron Blue and immobilized substrate onto Eupergit C, were important steps in the purification protocol. The electrophoretic mobility showed by the purified enzyme, agreed with a molecular mass of 14 kDa. The PLA2 activity against liposomes was determined by measurement of apparent absorbance changes at 340 nm. Also, high performance thin layer chromatography (HPTLC) analysis confirmed selective hydrolysis at the sn-2 position of soybean phospholipids. PLA2 exhibited millimolar calcium dependence and had slightly alkaline pH optimum. The enzyme was stimulated by auxins and completely inactivated by ammonium sulfate. Furthermore, it was irreversibly inactivated by p-bromo-phenacyl bromide, the specific inhibitor of secretory PLA2s. In addition, Glycine max PLA2 showed high stability against heat treatment and organic solvents. The enzyme showed activity toward multilamellar vesicles of soybean phospholipids, with a Vmax = 950 U/mg and a KM = 0.78 mM. The hyperbolic behavior observed was coherent with a hopping mode of action, one of the two characteristic interfacial mechanisms of PLA2s. All these data agree with the expected properties for a secretory PLA2 being the first soybean enzyme of this type. The new PLA2 developed lipolytic activity on a water in oil microemulsion reaction system, which is suitable for lysophospholipid production. These lysoderivatives are valuable biosurfactants for food and pharmaceutical industries.Fil: Minchiotti, Mariana Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; ArgentinaFil: Scalambro, Maria Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba. Facultad de Cs.agropecuarias. Departamento de Fundamentación Biologica; ArgentinaFil: Vargas, Laura Inés. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; ArgentinaFil: Coronel, Carlos Enrique. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Madoery, Ricardo Roman. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; ArgentinaElsevier Science Inc2008-04-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/58410Minchiotti, Mariana Cecilia; Scalambro, Maria Belen; Vargas, Laura Inés; Coronel, Carlos Enrique; Madoery, Ricardo Roman; Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties; Elsevier Science Inc; Enzyme and Microbial Technology; 42; 5; 4-4-2008; 389-3940141-0229CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0141022907003614info:eu-repo/semantics/altIdentifier/doi/10.1016/j.enzmictec.2007.11.015info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:42:53Zoai:ri.conicet.gov.ar:11336/58410instacron: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:34982025-10-22 11:42:53.392CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties
title Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties
spellingShingle Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties
Minchiotti, Mariana Cecilia
Phospholipase A2
Properties
Purification
Soybean
title_short Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties
title_full Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties
title_fullStr Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties
title_full_unstemmed Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties
title_sort Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties
dc.creator.none.fl_str_mv Minchiotti, Mariana Cecilia
Scalambro, Maria Belen
Vargas, Laura Inés
Coronel, Carlos Enrique
Madoery, Ricardo Roman
author Minchiotti, Mariana Cecilia
author_facet Minchiotti, Mariana Cecilia
Scalambro, Maria Belen
Vargas, Laura Inés
Coronel, Carlos Enrique
Madoery, Ricardo Roman
author_role author
author2 Scalambro, Maria Belen
Vargas, Laura Inés
Coronel, Carlos Enrique
Madoery, Ricardo Roman
author2_role author
author
author
author
dc.subject.none.fl_str_mv Phospholipase A2
Properties
Purification
Soybean
topic Phospholipase A2
Properties
Purification
Soybean
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv A protein with phospholipase A2 (PLA2) activity was isolated from soybean (Glycine max) seeds. The affinity column chromatography techniques by using Cibacron Blue and immobilized substrate onto Eupergit C, were important steps in the purification protocol. The electrophoretic mobility showed by the purified enzyme, agreed with a molecular mass of 14 kDa. The PLA2 activity against liposomes was determined by measurement of apparent absorbance changes at 340 nm. Also, high performance thin layer chromatography (HPTLC) analysis confirmed selective hydrolysis at the sn-2 position of soybean phospholipids. PLA2 exhibited millimolar calcium dependence and had slightly alkaline pH optimum. The enzyme was stimulated by auxins and completely inactivated by ammonium sulfate. Furthermore, it was irreversibly inactivated by p-bromo-phenacyl bromide, the specific inhibitor of secretory PLA2s. In addition, Glycine max PLA2 showed high stability against heat treatment and organic solvents. The enzyme showed activity toward multilamellar vesicles of soybean phospholipids, with a Vmax = 950 U/mg and a KM = 0.78 mM. The hyperbolic behavior observed was coherent with a hopping mode of action, one of the two characteristic interfacial mechanisms of PLA2s. All these data agree with the expected properties for a secretory PLA2 being the first soybean enzyme of this type. The new PLA2 developed lipolytic activity on a water in oil microemulsion reaction system, which is suitable for lysophospholipid production. These lysoderivatives are valuable biosurfactants for food and pharmaceutical industries.
Fil: Minchiotti, Mariana Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; Argentina
Fil: Scalambro, Maria Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba. Facultad de Cs.agropecuarias. Departamento de Fundamentación Biologica; Argentina
Fil: Vargas, Laura Inés. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; Argentina
Fil: Coronel, Carlos Enrique. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
Fil: Madoery, Ricardo Roman. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; Argentina
description A protein with phospholipase A2 (PLA2) activity was isolated from soybean (Glycine max) seeds. The affinity column chromatography techniques by using Cibacron Blue and immobilized substrate onto Eupergit C, were important steps in the purification protocol. The electrophoretic mobility showed by the purified enzyme, agreed with a molecular mass of 14 kDa. The PLA2 activity against liposomes was determined by measurement of apparent absorbance changes at 340 nm. Also, high performance thin layer chromatography (HPTLC) analysis confirmed selective hydrolysis at the sn-2 position of soybean phospholipids. PLA2 exhibited millimolar calcium dependence and had slightly alkaline pH optimum. The enzyme was stimulated by auxins and completely inactivated by ammonium sulfate. Furthermore, it was irreversibly inactivated by p-bromo-phenacyl bromide, the specific inhibitor of secretory PLA2s. In addition, Glycine max PLA2 showed high stability against heat treatment and organic solvents. The enzyme showed activity toward multilamellar vesicles of soybean phospholipids, with a Vmax = 950 U/mg and a KM = 0.78 mM. The hyperbolic behavior observed was coherent with a hopping mode of action, one of the two characteristic interfacial mechanisms of PLA2s. All these data agree with the expected properties for a secretory PLA2 being the first soybean enzyme of this type. The new PLA2 developed lipolytic activity on a water in oil microemulsion reaction system, which is suitable for lysophospholipid production. These lysoderivatives are valuable biosurfactants for food and pharmaceutical industries.
publishDate 2008
dc.date.none.fl_str_mv 2008-04-04
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/11336/58410
Minchiotti, Mariana Cecilia; Scalambro, Maria Belen; Vargas, Laura Inés; Coronel, Carlos Enrique; Madoery, Ricardo Roman; Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties; Elsevier Science Inc; Enzyme and Microbial Technology; 42; 5; 4-4-2008; 389-394
0141-0229
CONICET Digital
CONICET
url http://hdl.handle.net/11336/58410
identifier_str_mv Minchiotti, Mariana Cecilia; Scalambro, Maria Belen; Vargas, Laura Inés; Coronel, Carlos Enrique; Madoery, Ricardo Roman; Isolation of phospholipase A2 from soybean (Glycine max) seeds: The study of its enzymatic properties; Elsevier Science Inc; Enzyme and Microbial Technology; 42; 5; 4-4-2008; 389-394
0141-0229
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0141022907003614
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.enzmictec.2007.11.015
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science Inc
publisher.none.fl_str_mv Elsevier Science Inc
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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