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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/58410
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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 Técnicas2025-09-29T10:12:58Zoai: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:34982025-09-29 10:12:59.129CONICET 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 |
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reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
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CONICET Digital (CONICET) |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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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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1844614041930039296 |
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13.070432 |