Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption

Autores
Boeris, Valeria; Balce, Izabella; Vennapusa, Rami Reddy; Rodríguez, Miguel Arévalo; Picó, Guillermo Alfredo; Lahore, Marcelo Fernández
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
β-Galactosidase is a hydrolase enzyme that catalyzes the hydrolysis of β-galactosides into monosaccharides; its major application in the food industry is to reduce the content of lactose in lactic products. The aim of this work is to recover this enzyme from a cell lysate by adsorption onto Streamline-DEAE in an expanded bed, avoiding, as much as possible, biomass deposition onto the adsorbent matrix. So as to achieve less cell debris-matrix interaction, the adsorbent surface was covered with polyvinyl pyrrolidone. The enzyme showed to bind in the same extent to naked and covered Streamline-DEAE (65. mg β-gal/g matrix) in batch mode in the absence of any biomass. The kinetics of the adsorption process was studied and no effect of the polyvinyl pyrrolidone covering was found. The optimal conditions for the recovery were achieved by using a lysate made of 40% wet weight of cells, a polyvinyl pyrrolidone-covered matrix/lysate ratio of 10% and carrying out the adsorption process in expanded bed with recirculation over 2. h in 20. mM phosphate buffer pH 7.4. The fraction recovered after the elution contained 65% of the initial amount of enzyme with a 12.6-fold increased specific activity with respect to the lysate. The polyvinyl pyrrolidone content in the eluate was determined and found negligible. The remarkable point of this work is that it was possible to partially purify the enzyme using a feedstock containing an unusually high biomass concentration in the presence of polyvinyl pyrrolidone onto weak anion exchangers.
Fil: Boeris, Valeria. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina
Fil: Balce, Izabella. Universitat Bremen. School of Engineering and Science Jacobs; Alemania
Fil: Vennapusa, Rami Reddy. Universitat Bremen. School of Engineering and Science Jacobs; Alemania
Fil: Rodríguez, Miguel Arévalo. Universidad Pablo de Olavide; España
Fil: Picó, Guillermo Alfredo. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina
Fil: Lahore, Marcelo Fernández. Universitat Bremen. School of Engineering and Science Jacobs; Alemania
Materia
Β-GAL
Β-GALACTOSIDASE
EBA
EXPANDED BED ADSORPTION
PVP
STREAMLINE-DEAE
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:11336/139025
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/139025
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorptionBoeris, ValeriaBalce, IzabellaVennapusa, Rami ReddyRodríguez, Miguel ArévaloPicó, Guillermo AlfredoLahore, Marcelo FernándezΒ-GALΒ-GALACTOSIDASEEBAEXPANDED BED ADSORPTIONPVPSTREAMLINE-DEAEhttps://purl.org/becyt/ford/2.9https://purl.org/becyt/ford/2β-Galactosidase is a hydrolase enzyme that catalyzes the hydrolysis of β-galactosides into monosaccharides; its major application in the food industry is to reduce the content of lactose in lactic products. The aim of this work is to recover this enzyme from a cell lysate by adsorption onto Streamline-DEAE in an expanded bed, avoiding, as much as possible, biomass deposition onto the adsorbent matrix. So as to achieve less cell debris-matrix interaction, the adsorbent surface was covered with polyvinyl pyrrolidone. The enzyme showed to bind in the same extent to naked and covered Streamline-DEAE (65. mg β-gal/g matrix) in batch mode in the absence of any biomass. The kinetics of the adsorption process was studied and no effect of the polyvinyl pyrrolidone covering was found. The optimal conditions for the recovery were achieved by using a lysate made of 40% wet weight of cells, a polyvinyl pyrrolidone-covered matrix/lysate ratio of 10% and carrying out the adsorption process in expanded bed with recirculation over 2. h in 20. mM phosphate buffer pH 7.4. The fraction recovered after the elution contained 65% of the initial amount of enzyme with a 12.6-fold increased specific activity with respect to the lysate. The polyvinyl pyrrolidone content in the eluate was determined and found negligible. The remarkable point of this work is that it was possible to partially purify the enzyme using a feedstock containing an unusually high biomass concentration in the presence of polyvinyl pyrrolidone onto weak anion exchangers.Fil: Boeris, Valeria. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Balce, Izabella. Universitat Bremen. School of Engineering and Science Jacobs; AlemaniaFil: Vennapusa, Rami Reddy. Universitat Bremen. School of Engineering and Science Jacobs; AlemaniaFil: Rodríguez, Miguel Arévalo. Universidad Pablo de Olavide; EspañaFil: Picó, Guillermo Alfredo. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Lahore, Marcelo Fernández. Universitat Bremen. School of Engineering and Science Jacobs; AlemaniaElsevier Science2012-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/139025Boeris, Valeria; Balce, Izabella; Vennapusa, Rami Reddy; Rodríguez, Miguel Arévalo; Picó, Guillermo Alfredo; et al.; Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption; Elsevier Science; Journal of Chromatography B; 900; 7-2012; 32-371570-0232CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S1570023212003054info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jchromb.2012.05.024info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:29:26Zoai:ri.conicet.gov.ar:11336/139025instacron: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 10:29:26.965CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption
title Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption
spellingShingle Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption
Boeris, Valeria
Β-GAL
Β-GALACTOSIDASE
EBA
EXPANDED BED ADSORPTION
PVP
STREAMLINE-DEAE
title_short Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption
title_full Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption
title_fullStr Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption
title_full_unstemmed Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption
title_sort Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption
dc.creator.none.fl_str_mv Boeris, Valeria
Balce, Izabella
Vennapusa, Rami Reddy
Rodríguez, Miguel Arévalo
Picó, Guillermo Alfredo
Lahore, Marcelo Fernández
author Boeris, Valeria
author_facet Boeris, Valeria
Balce, Izabella
Vennapusa, Rami Reddy
Rodríguez, Miguel Arévalo
Picó, Guillermo Alfredo
Lahore, Marcelo Fernández
author_role author
author2 Balce, Izabella
Vennapusa, Rami Reddy
Rodríguez, Miguel Arévalo
Picó, Guillermo Alfredo
Lahore, Marcelo Fernández
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Β-GAL
Β-GALACTOSIDASE
EBA
EXPANDED BED ADSORPTION
PVP
STREAMLINE-DEAE
topic Β-GAL
Β-GALACTOSIDASE
EBA
EXPANDED BED ADSORPTION
PVP
STREAMLINE-DEAE
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.9
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv β-Galactosidase is a hydrolase enzyme that catalyzes the hydrolysis of β-galactosides into monosaccharides; its major application in the food industry is to reduce the content of lactose in lactic products. The aim of this work is to recover this enzyme from a cell lysate by adsorption onto Streamline-DEAE in an expanded bed, avoiding, as much as possible, biomass deposition onto the adsorbent matrix. So as to achieve less cell debris-matrix interaction, the adsorbent surface was covered with polyvinyl pyrrolidone. The enzyme showed to bind in the same extent to naked and covered Streamline-DEAE (65. mg β-gal/g matrix) in batch mode in the absence of any biomass. The kinetics of the adsorption process was studied and no effect of the polyvinyl pyrrolidone covering was found. The optimal conditions for the recovery were achieved by using a lysate made of 40% wet weight of cells, a polyvinyl pyrrolidone-covered matrix/lysate ratio of 10% and carrying out the adsorption process in expanded bed with recirculation over 2. h in 20. mM phosphate buffer pH 7.4. The fraction recovered after the elution contained 65% of the initial amount of enzyme with a 12.6-fold increased specific activity with respect to the lysate. The polyvinyl pyrrolidone content in the eluate was determined and found negligible. The remarkable point of this work is that it was possible to partially purify the enzyme using a feedstock containing an unusually high biomass concentration in the presence of polyvinyl pyrrolidone onto weak anion exchangers.
Fil: Boeris, Valeria. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina
Fil: Balce, Izabella. Universitat Bremen. School of Engineering and Science Jacobs; Alemania
Fil: Vennapusa, Rami Reddy. Universitat Bremen. School of Engineering and Science Jacobs; Alemania
Fil: Rodríguez, Miguel Arévalo. Universidad Pablo de Olavide; España
Fil: Picó, Guillermo Alfredo. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; Argentina
Fil: Lahore, Marcelo Fernández. Universitat Bremen. School of Engineering and Science Jacobs; Alemania
description β-Galactosidase is a hydrolase enzyme that catalyzes the hydrolysis of β-galactosides into monosaccharides; its major application in the food industry is to reduce the content of lactose in lactic products. The aim of this work is to recover this enzyme from a cell lysate by adsorption onto Streamline-DEAE in an expanded bed, avoiding, as much as possible, biomass deposition onto the adsorbent matrix. So as to achieve less cell debris-matrix interaction, the adsorbent surface was covered with polyvinyl pyrrolidone. The enzyme showed to bind in the same extent to naked and covered Streamline-DEAE (65. mg β-gal/g matrix) in batch mode in the absence of any biomass. The kinetics of the adsorption process was studied and no effect of the polyvinyl pyrrolidone covering was found. The optimal conditions for the recovery were achieved by using a lysate made of 40% wet weight of cells, a polyvinyl pyrrolidone-covered matrix/lysate ratio of 10% and carrying out the adsorption process in expanded bed with recirculation over 2. h in 20. mM phosphate buffer pH 7.4. The fraction recovered after the elution contained 65% of the initial amount of enzyme with a 12.6-fold increased specific activity with respect to the lysate. The polyvinyl pyrrolidone content in the eluate was determined and found negligible. The remarkable point of this work is that it was possible to partially purify the enzyme using a feedstock containing an unusually high biomass concentration in the presence of polyvinyl pyrrolidone onto weak anion exchangers.
publishDate 2012
dc.date.none.fl_str_mv 2012-07
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/11336/139025
Boeris, Valeria; Balce, Izabella; Vennapusa, Rami Reddy; Rodríguez, Miguel Arévalo; Picó, Guillermo Alfredo; et al.; Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption; Elsevier Science; Journal of Chromatography B; 900; 7-2012; 32-37
1570-0232
CONICET Digital
CONICET
url http://hdl.handle.net/11336/139025
identifier_str_mv Boeris, Valeria; Balce, Izabella; Vennapusa, Rami Reddy; Rodríguez, Miguel Arévalo; Picó, Guillermo Alfredo; et al.; Production, recovery and purification of a recombinant β-galactosidase by expanded bed anion exchange adsorption; Elsevier Science; Journal of Chromatography B; 900; 7-2012; 32-37
1570-0232
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/abs/pii/S1570023212003054
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jchromb.2012.05.024
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
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
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 13.070432

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