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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/41024
Ver los metadatos del registro completo
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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 Técnicas2025-09-29T09:43:37Zoai: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:34982025-09-29 09:43:37.927CONICET 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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1844613373455499264 |
score |
13.070432 |