Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization

Autores
Rivero, Cintia Wanda; de Benedetti, Eliana Celeste; Lopez Gallego, F; Pessela, B. C.; Guisán, J. M.; Trelles, Jorge Abel
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Ribavirin is a synthetic guanosine analogue with a broad‐spectrum of antiviral activity. It is clinically
effective against several viruses, such as respiratory syncytial virus, several hemorrhagic fever viruses and HCV when combined with pegylated interferon‐. Phosphopentomutase (PPM) catalyzes the transfer of intramolecular phosphate (from C1 to C5) on ribose, and is involved in pentose phosphate pathway and in purine metabolism.  Reactions catalyzed by this enzyme are useful for nucleoside analogues production. However, out of its natural environment PPM is unstable and its stability is affected by parameters such as pH and temperature. Therefore, to irreversibly immobilize this enzyme, it needs to be stabilized. In this work, PPM from Escherichia coli ATCC 4157 was overexpressed, purified, stabilized at alkaline pH and immobilized on several supports. The activity of different additives as stabilizing agents was evaluated, and the best result was found using 10% (v/v) glycerol. Under this condition, PPM maintained 86% of its initial activity at pH 10 after 18 h incubation, which allowed further covalent immobilization of this enzyme on glyoxyl‐agarose with a high yield. 
Fil: Rivero, Cintia Wanda. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Biotecnología Sustentable; Argentina
Fil: de Benedetti, Eliana Celeste. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Biotecnología Sustentable; Argentina
Fil: Lopez Gallego, F. Heterogeneous Biocatalysis Group; España. Basque Foundation for Science; España
Fil: Pessela, B. C.. Research Institute of Food Science; España. Polytechnic Institute of Science and Technology; Angola
Fil: Guisán, J. M.. Institute of Catalysis and Petrochemistry; España
Fil: Trelles, Jorge Abel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Biotecnología Sustentable; Argentina
Materia
Stabilizing Agents
Glyoxyl-Agarose
Ribavirin
Escherichia Coli
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:11336/41024
Acceder
id CONICETDig_f5ac8813887ba7107d616344e61fa806
oai_identifier_str oai:ri.conicet.gov.ar:11336/41024
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilizationRivero, Cintia Wandade Benedetti, Eliana CelesteLopez Gallego, FPessela, B. C.Guisán, J. M.Trelles, Jorge AbelStabilizing AgentsGlyoxyl-AgaroseRibavirinEscherichia Colihttps://purl.org/becyt/ford/2.9https://purl.org/becyt/ford/2https://purl.org/becyt/ford/2.9https://purl.org/becyt/ford/2Ribavirin is a synthetic guanosine analogue with a broad‐spectrum of antiviral activity. It is clinically <br />effective against several viruses, such as respiratory syncytial virus, several hemorrhagic fever viruses and HCV when combined with pegylated interferon‐. Phosphopentomutase (PPM) catalyzes the transfer of intramolecular phosphate (from C1 to C5) on ribose, and is involved in pentose phosphate pathway and in purine metabolism.  Reactions catalyzed by this enzyme are useful for nucleoside analogues production. However, out of its natural environment PPM is unstable and its stability is affected by parameters such as pH and temperature. Therefore, to irreversibly immobilize this enzyme, it needs to be stabilized. In this work, PPM from Escherichia coli ATCC 4157 was overexpressed, purified, stabilized at alkaline pH and immobilized on several supports. The activity of different additives as stabilizing agents was evaluated, and the best result was found using 10% (v/v) glycerol. Under this condition, PPM maintained 86% of its initial activity at pH 10 after 18 h incubation, which allowed further covalent immobilization of this enzyme on glyoxyl‐agarose with a high yield. Fil: Rivero, Cintia Wanda. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Biotecnología Sustentable; ArgentinaFil: de Benedetti, Eliana Celeste. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Biotecnología Sustentable; ArgentinaFil: Lopez Gallego, F. Heterogeneous Biocatalysis Group; España. Basque Foundation for Science; EspañaFil: Pessela, B. C.. Research Institute of Food Science; España. Polytechnic Institute of Science and Technology; AngolaFil: Guisán, J. M.. Institute of Catalysis and Petrochemistry; EspañaFil: Trelles, Jorge Abel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Biotecnología Sustentable; ArgentinaElsevier Science2017-03info: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/11336/41024Rivero, Cintia Wanda; de Benedetti, Eliana Celeste; Lopez Gallego, F; Pessela, B. C.; Guisán, J. M.; et al.; Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization; Elsevier Science; Journal of Biotechnology; 249; 3-2017; 34-410168-1656CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.jbiotec.2017.03.027info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0168165617301323info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:36:57Zoai:ri.conicet.gov.ar:11336/41024instacron: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:34982025-10-15 14:36:57.525CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization
title Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization
spellingShingle Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization
Rivero, Cintia Wanda
Stabilizing Agents
Glyoxyl-Agarose
Ribavirin
Escherichia Coli
title_short Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization
title_full Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization
title_fullStr Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization
title_full_unstemmed Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization
title_sort Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization
dc.creator.none.fl_str_mv Rivero, Cintia Wanda
de Benedetti, Eliana Celeste
Lopez Gallego, F
Pessela, B. C.
Guisán, J. M.
Trelles, Jorge Abel
author Rivero, Cintia Wanda
author_facet Rivero, Cintia Wanda
de Benedetti, Eliana Celeste
Lopez Gallego, F
Pessela, B. C.
Guisán, J. M.
Trelles, Jorge Abel
author_role author
author2 de Benedetti, Eliana Celeste
Lopez Gallego, F
Pessela, B. C.
Guisán, J. M.
Trelles, Jorge Abel
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Stabilizing Agents
Glyoxyl-Agarose
Ribavirin
Escherichia Coli
topic Stabilizing Agents
Glyoxyl-Agarose
Ribavirin
Escherichia Coli
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.9
https://purl.org/becyt/ford/2
https://purl.org/becyt/ford/2.9
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv Ribavirin is a synthetic guanosine analogue with a broad‐spectrum of antiviral activity. It is clinically <br />effective against several viruses, such as respiratory syncytial virus, several hemorrhagic fever viruses and HCV when combined with pegylated interferon‐. Phosphopentomutase (PPM) catalyzes the transfer of intramolecular phosphate (from C1 to C5) on ribose, and is involved in pentose phosphate pathway and in purine metabolism.  Reactions catalyzed by this enzyme are useful for nucleoside analogues production. However, out of its natural environment PPM is unstable and its stability is affected by parameters such as pH and temperature. Therefore, to irreversibly immobilize this enzyme, it needs to be stabilized. In this work, PPM from Escherichia coli ATCC 4157 was overexpressed, purified, stabilized at alkaline pH and immobilized on several supports. The activity of different additives as stabilizing agents was evaluated, and the best result was found using 10% (v/v) glycerol. Under this condition, PPM maintained 86% of its initial activity at pH 10 after 18 h incubation, which allowed further covalent immobilization of this enzyme on glyoxyl‐agarose with a high yield. 
Fil: Rivero, Cintia Wanda. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Biotecnología Sustentable; Argentina
Fil: de Benedetti, Eliana Celeste. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Biotecnología Sustentable; Argentina
Fil: Lopez Gallego, F. Heterogeneous Biocatalysis Group; España. Basque Foundation for Science; España
Fil: Pessela, B. C.. Research Institute of Food Science; España. Polytechnic Institute of Science and Technology; Angola
Fil: Guisán, J. M.. Institute of Catalysis and Petrochemistry; España
Fil: Trelles, Jorge Abel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Biotecnología Sustentable; Argentina
description Ribavirin is a synthetic guanosine analogue with a broad‐spectrum of antiviral activity. It is clinically <br />effective against several viruses, such as respiratory syncytial virus, several hemorrhagic fever viruses and HCV when combined with pegylated interferon‐. Phosphopentomutase (PPM) catalyzes the transfer of intramolecular phosphate (from C1 to C5) on ribose, and is involved in pentose phosphate pathway and in purine metabolism.  Reactions catalyzed by this enzyme are useful for nucleoside analogues production. However, out of its natural environment PPM is unstable and its stability is affected by parameters such as pH and temperature. Therefore, to irreversibly immobilize this enzyme, it needs to be stabilized. In this work, PPM from Escherichia coli ATCC 4157 was overexpressed, purified, stabilized at alkaline pH and immobilized on several supports. The activity of different additives as stabilizing agents was evaluated, and the best result was found using 10% (v/v) glycerol. Under this condition, PPM maintained 86% of its initial activity at pH 10 after 18 h incubation, which allowed further covalent immobilization of this enzyme on glyoxyl‐agarose with a high yield. 
publishDate 2017
dc.date.none.fl_str_mv 2017-03
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/11336/41024
Rivero, Cintia Wanda; de Benedetti, Eliana Celeste; Lopez Gallego, F; Pessela, B. C.; Guisán, J. M.; et al.; Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization; Elsevier Science; Journal of Biotechnology; 249; 3-2017; 34-41
0168-1656
CONICET Digital
CONICET
url http://hdl.handle.net/11336/41024
identifier_str_mv Rivero, Cintia Wanda; de Benedetti, Eliana Celeste; Lopez Gallego, F; Pessela, B. C.; Guisán, J. M.; et al.; Biosynthesis of an antiviral compound using a stabilized phosphopentomutase by multipoint covalent immobilization; Elsevier Science; Journal of Biotechnology; 249; 3-2017; 34-41
0168-1656
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jbiotec.2017.03.027
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0168165617301323
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
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 13.22299

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