His-tag presence modulates enzymatic activity both in solution and at lipid interfaces

Autores
Flores, Sandra S.; Perillo, Maria Angelica; Nolan, María Verónica; Sanchez, Julieta Maria
Año de publicación
2021
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
β-Galactosidase (β-Gal) is an important biotechnological enzyme used in the dairy industry, pharmacology and in molecular biology. This enzyme has a commercial application for lactose hydrolysis in dairy products. Milk processing with β-Gal before milk is commercialized is important to solve nutritional (lactose intolerance) and technological (crystallization of dairy products) problems. In this context, it is important that the activity of β-Gal be evaluated in heterogeneous media.β-Galactosidase (β-Gal) is an important biotechnological enzyme used in the dairy industry, pharmacology and in molecular biology. This enzyme has a commercial application for lactose hydrolysis in dairy products. Milk processing with β-Gal before milk is commercialized is important to solve nutritional (lactose intolerance) and technological (crystallization of dairy products) problems. In this context, it is important that the activity of β-Gal be evaluated in heterogeneous media. In our laboratory we have overexpressed a recombinant β-galactosidase in Escherichia coli (E. coli). This enzyme differs from its native version (β-GalWT) in that 6 histidine residues have been added to the carboxyl terminus in the primary sequence (β-GalHis), which allows its purification by immobilized metal affinity chromatography (IMAC). In this work we compared the functionality of both proteins and evaluated their catalytic behavior on the kinetics of lactose hydrolysis. We observed a significant reduction in the enzymatic activity of β-GalHis with respect to β-GalWT. Our studies also focus in studying the activity of both β-Gals in the presence of multilamellar vesicles (MLVs) of different composition. We conclude that the additional positive charges β-GalHis (belonging from histidine residues) promotes the interaction of the protein with negatively charged interfaces favoring the effect shown against neutral interfaces.
Fil: Flores, Sandra S.. 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. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina
Fil: Perillo, Maria Angelica. 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. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina
Fil: Nolan, María Verónica. 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. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina
Fil: Sanchez, Julieta Maria. 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. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina
XLIX Reunión Anual Sociedad Argentina de Biofísica
Buenos Aires
Argentina
Sociedad Argentina de Biofisica
Materia
BETA GALACTOSIDASE
ENZYMATIC ACTIVITY
INTERFACIAL ACTIVITY
BETA GAL HIS
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:11336/189162
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/189162
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling His-tag presence modulates enzymatic activity both in solution and at lipid interfacesFlores, Sandra S.Perillo, Maria AngelicaNolan, María VerónicaSanchez, Julieta MariaBETA GALACTOSIDASEENZYMATIC ACTIVITYINTERFACIAL ACTIVITYBETA GAL HIShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1β-Galactosidase (β-Gal) is an important biotechnological enzyme used in the dairy industry, pharmacology and in molecular biology. This enzyme has a commercial application for lactose hydrolysis in dairy products. Milk processing with β-Gal before milk is commercialized is important to solve nutritional (lactose intolerance) and technological (crystallization of dairy products) problems. In this context, it is important that the activity of β-Gal be evaluated in heterogeneous media.β-Galactosidase (β-Gal) is an important biotechnological enzyme used in the dairy industry, pharmacology and in molecular biology. This enzyme has a commercial application for lactose hydrolysis in dairy products. Milk processing with β-Gal before milk is commercialized is important to solve nutritional (lactose intolerance) and technological (crystallization of dairy products) problems. In this context, it is important that the activity of β-Gal be evaluated in heterogeneous media. In our laboratory we have overexpressed a recombinant β-galactosidase in Escherichia coli (E. coli). This enzyme differs from its native version (β-GalWT) in that 6 histidine residues have been added to the carboxyl terminus in the primary sequence (β-GalHis), which allows its purification by immobilized metal affinity chromatography (IMAC). In this work we compared the functionality of both proteins and evaluated their catalytic behavior on the kinetics of lactose hydrolysis. We observed a significant reduction in the enzymatic activity of β-GalHis with respect to β-GalWT. Our studies also focus in studying the activity of both β-Gals in the presence of multilamellar vesicles (MLVs) of different composition. We conclude that the additional positive charges β-GalHis (belonging from histidine residues) promotes the interaction of the protein with negatively charged interfaces favoring the effect shown against neutral interfaces.Fil: Flores, Sandra S.. 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. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; ArgentinaFil: Perillo, Maria Angelica. 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. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; ArgentinaFil: Nolan, María Verónica. 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. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; ArgentinaFil: Sanchez, Julieta Maria. 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. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; ArgentinaXLIX Reunión Anual Sociedad Argentina de BiofísicaBuenos AiresArgentinaSociedad Argentina de BiofisicaSociedad Argentina de BiofísicaDelfino, Jose MariaCelej, Maria SoledadMangialavori, Irene CeciliaAcierno, Juan2021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/189162His-tag presence modulates enzymatic activity both in solution and at lipid interfaces; XLIX Reunión Anual Sociedad Argentina de Biofísica; Buenos Aires; Argentina; 2021; 112-112978-987-27591-9-3CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://biofisica.org.ar/reuniones-cientificas/reunionsab-previas/Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:24:42Zoai:ri.conicet.gov.ar:11336/189162instacron: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 10:24:43.209CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv His-tag presence modulates enzymatic activity both in solution and at lipid interfaces
title His-tag presence modulates enzymatic activity both in solution and at lipid interfaces
spellingShingle His-tag presence modulates enzymatic activity both in solution and at lipid interfaces
Flores, Sandra S.
BETA GALACTOSIDASE
ENZYMATIC ACTIVITY
INTERFACIAL ACTIVITY
BETA GAL HIS
title_short His-tag presence modulates enzymatic activity both in solution and at lipid interfaces
title_full His-tag presence modulates enzymatic activity both in solution and at lipid interfaces
title_fullStr His-tag presence modulates enzymatic activity both in solution and at lipid interfaces
title_full_unstemmed His-tag presence modulates enzymatic activity both in solution and at lipid interfaces
title_sort His-tag presence modulates enzymatic activity both in solution and at lipid interfaces
dc.creator.none.fl_str_mv Flores, Sandra S.
Perillo, Maria Angelica
Nolan, María Verónica
Sanchez, Julieta Maria
author Flores, Sandra S.
author_facet Flores, Sandra S.
Perillo, Maria Angelica
Nolan, María Verónica
Sanchez, Julieta Maria
author_role author
author2 Perillo, Maria Angelica
Nolan, María Verónica
Sanchez, Julieta Maria
author2_role author
author
author
dc.contributor.none.fl_str_mv Delfino, Jose Maria
Celej, Maria Soledad
Mangialavori, Irene Cecilia
Acierno, Juan
dc.subject.none.fl_str_mv BETA GALACTOSIDASE
ENZYMATIC ACTIVITY
INTERFACIAL ACTIVITY
BETA GAL HIS
topic BETA GALACTOSIDASE
ENZYMATIC ACTIVITY
INTERFACIAL ACTIVITY
BETA GAL HIS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv β-Galactosidase (β-Gal) is an important biotechnological enzyme used in the dairy industry, pharmacology and in molecular biology. This enzyme has a commercial application for lactose hydrolysis in dairy products. Milk processing with β-Gal before milk is commercialized is important to solve nutritional (lactose intolerance) and technological (crystallization of dairy products) problems. In this context, it is important that the activity of β-Gal be evaluated in heterogeneous media.β-Galactosidase (β-Gal) is an important biotechnological enzyme used in the dairy industry, pharmacology and in molecular biology. This enzyme has a commercial application for lactose hydrolysis in dairy products. Milk processing with β-Gal before milk is commercialized is important to solve nutritional (lactose intolerance) and technological (crystallization of dairy products) problems. In this context, it is important that the activity of β-Gal be evaluated in heterogeneous media. In our laboratory we have overexpressed a recombinant β-galactosidase in Escherichia coli (E. coli). This enzyme differs from its native version (β-GalWT) in that 6 histidine residues have been added to the carboxyl terminus in the primary sequence (β-GalHis), which allows its purification by immobilized metal affinity chromatography (IMAC). In this work we compared the functionality of both proteins and evaluated their catalytic behavior on the kinetics of lactose hydrolysis. We observed a significant reduction in the enzymatic activity of β-GalHis with respect to β-GalWT. Our studies also focus in studying the activity of both β-Gals in the presence of multilamellar vesicles (MLVs) of different composition. We conclude that the additional positive charges β-GalHis (belonging from histidine residues) promotes the interaction of the protein with negatively charged interfaces favoring the effect shown against neutral interfaces.
Fil: Flores, Sandra S.. 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. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina
Fil: Perillo, Maria Angelica. 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. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina
Fil: Nolan, María Verónica. 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. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina
Fil: Sanchez, Julieta Maria. 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. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina
XLIX Reunión Anual Sociedad Argentina de Biofísica
Buenos Aires
Argentina
Sociedad Argentina de Biofisica
description β-Galactosidase (β-Gal) is an important biotechnological enzyme used in the dairy industry, pharmacology and in molecular biology. This enzyme has a commercial application for lactose hydrolysis in dairy products. Milk processing with β-Gal before milk is commercialized is important to solve nutritional (lactose intolerance) and technological (crystallization of dairy products) problems. In this context, it is important that the activity of β-Gal be evaluated in heterogeneous media.β-Galactosidase (β-Gal) is an important biotechnological enzyme used in the dairy industry, pharmacology and in molecular biology. This enzyme has a commercial application for lactose hydrolysis in dairy products. Milk processing with β-Gal before milk is commercialized is important to solve nutritional (lactose intolerance) and technological (crystallization of dairy products) problems. In this context, it is important that the activity of β-Gal be evaluated in heterogeneous media. In our laboratory we have overexpressed a recombinant β-galactosidase in Escherichia coli (E. coli). This enzyme differs from its native version (β-GalWT) in that 6 histidine residues have been added to the carboxyl terminus in the primary sequence (β-GalHis), which allows its purification by immobilized metal affinity chromatography (IMAC). In this work we compared the functionality of both proteins and evaluated their catalytic behavior on the kinetics of lactose hydrolysis. We observed a significant reduction in the enzymatic activity of β-GalHis with respect to β-GalWT. Our studies also focus in studying the activity of both β-Gals in the presence of multilamellar vesicles (MLVs) of different composition. We conclude that the additional positive charges β-GalHis (belonging from histidine residues) promotes the interaction of the protein with negatively charged interfaces favoring the effect shown against neutral interfaces.
publishDate 2021
dc.date.none.fl_str_mv 2021
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/189162
His-tag presence modulates enzymatic activity both in solution and at lipid interfaces; XLIX Reunión Anual Sociedad Argentina de Biofísica; Buenos Aires; Argentina; 2021; 112-112
978-987-27591-9-3
CONICET Digital
CONICET
url http://hdl.handle.net/11336/189162
identifier_str_mv His-tag presence modulates enzymatic activity both in solution and at lipid interfaces; XLIX Reunión Anual Sociedad Argentina de Biofísica; Buenos Aires; Argentina; 2021; 112-112
978-987-27591-9-3
CONICET Digital
CONICET
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language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://biofisica.org.ar/reuniones-cientificas/reunionsab-previas/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
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rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
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dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Sociedad Argentina de Biofísica
publisher.none.fl_str_mv Sociedad Argentina de Biofísica
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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reponame_str CONICET Digital (CONICET)
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