Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging time
- Autores
- Burgos, Martha Ines; Carrer, Dolores Catalina; Perillo, Maria Angelica
- Año de publicación
- 2013
- Idioma
- inglés
- Tipo de recurso
- documento de conferencia
- Estado
- versión publicada
- Descripción
- Protein encapsulation in solid matrixes is of interest for biotechnological purposes and it also serves as a model of molecular crowding. We have successfully entrapped the enzyme β-galactosidase (β-gal) in silicate gels via a sol-gel reaction. The catalytic activity of encapsulated β-gal (Eβ-gal) was compared with the activity of the soluble form (Sβ-gal) with two different substrates (ONPG and PNPG) and at different aging times. Data were analyzed with a Michaellis-Menten model. With the substrate ONPG, we found a Vmax value higher for Eβ-gal than for Sβ-gal and this difference increased at longer aging times. On the contrary, with PNPG, Vmax was not affected neither by the enzyme condition (Sβ-gal and Eβ-gal) nor by the aging time. The porous structure of the silicate matrix was also studied as a function of aging time of the gels. Thus, fluorescence correlation spectroscopy (FCS) was applied through the analysis of diffusional properties of fluorescent probes of different sizes, entrapped in the gel matrix. We observed an anomalous diffusion in some conditions, suggesting a self-similar pore structure, and aging time dependent changes in diffusion coefficients. Taken together our result supports the hypothesis that there is a relationship between the catalytic mechanism proposed for β-gal and water availability in the silicate gel since the ratio between bulk and structured water depends on the pore size.
Fil: Burgos, Martha Ines. 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: Carrer, Dolores Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; 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
XLII Reunión Anual de la Sociedad Argentina de Biofísica
Villa Carlos Paz
Argentina
Sociedad Argentina de Biofísica - Materia
-
Silicate matrix
Beta-galactosidase
P-nitrophenylphosphate
Fluorescencence correlation spectroscopy
Porous structure - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
.jpg)
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/237833
Ver los metadatos del registro completo
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Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging timeBurgos, Martha InesCarrer, Dolores CatalinaPerillo, Maria AngelicaSilicate matrixBeta-galactosidaseP-nitrophenylphosphateFluorescencence correlation spectroscopyPorous structurehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Protein encapsulation in solid matrixes is of interest for biotechnological purposes and it also serves as a model of molecular crowding. We have successfully entrapped the enzyme β-galactosidase (β-gal) in silicate gels via a sol-gel reaction. The catalytic activity of encapsulated β-gal (Eβ-gal) was compared with the activity of the soluble form (Sβ-gal) with two different substrates (ONPG and PNPG) and at different aging times. Data were analyzed with a Michaellis-Menten model. With the substrate ONPG, we found a Vmax value higher for Eβ-gal than for Sβ-gal and this difference increased at longer aging times. On the contrary, with PNPG, Vmax was not affected neither by the enzyme condition (Sβ-gal and Eβ-gal) nor by the aging time. The porous structure of the silicate matrix was also studied as a function of aging time of the gels. Thus, fluorescence correlation spectroscopy (FCS) was applied through the analysis of diffusional properties of fluorescent probes of different sizes, entrapped in the gel matrix. We observed an anomalous diffusion in some conditions, suggesting a self-similar pore structure, and aging time dependent changes in diffusion coefficients. Taken together our result supports the hypothesis that there is a relationship between the catalytic mechanism proposed for β-gal and water availability in the silicate gel since the ratio between bulk and structured water depends on the pore size.Fil: Burgos, Martha Ines. 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: Carrer, Dolores Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; 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; ArgentinaXLII Reunión Anual de la Sociedad Argentina de BiofísicaVilla Carlos PazArgentinaSociedad Argentina de BiofísicaSociedad Argentina de BiofísicaFanani, Maria LauraWilke, NataliaFidelio, Gerardo Daniel2013info: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/11336/237833Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging time; XLII Reunión Anual de la Sociedad Argentina de Biofísica; Villa Carlos Paz; Argentina; 2013; 59-59978-987-27591-2-4CONICET 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 Técnicas2025-10-29T11:40:25Zoai:ri.conicet.gov.ar:11336/237833instacron: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:34982025-10-29 11:40:25.475CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
| dc.title.none.fl_str_mv |
Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging time |
| title |
Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging time |
| spellingShingle |
Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging time Burgos, Martha Ines Silicate matrix Beta-galactosidase P-nitrophenylphosphate Fluorescencence correlation spectroscopy Porous structure |
| title_short |
Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging time |
| title_full |
Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging time |
| title_fullStr |
Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging time |
| title_full_unstemmed |
Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging time |
| title_sort |
Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging time |
| dc.creator.none.fl_str_mv |
Burgos, Martha Ines Carrer, Dolores Catalina Perillo, Maria Angelica |
| author |
Burgos, Martha Ines |
| author_facet |
Burgos, Martha Ines Carrer, Dolores Catalina Perillo, Maria Angelica |
| author_role |
author |
| author2 |
Carrer, Dolores Catalina Perillo, Maria Angelica |
| author2_role |
author author |
| dc.contributor.none.fl_str_mv |
Fanani, Maria Laura Wilke, Natalia Fidelio, Gerardo Daniel |
| dc.subject.none.fl_str_mv |
Silicate matrix Beta-galactosidase P-nitrophenylphosphate Fluorescencence correlation spectroscopy Porous structure |
| topic |
Silicate matrix Beta-galactosidase P-nitrophenylphosphate Fluorescencence correlation spectroscopy Porous structure |
| purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
| dc.description.none.fl_txt_mv |
Protein encapsulation in solid matrixes is of interest for biotechnological purposes and it also serves as a model of molecular crowding. We have successfully entrapped the enzyme β-galactosidase (β-gal) in silicate gels via a sol-gel reaction. The catalytic activity of encapsulated β-gal (Eβ-gal) was compared with the activity of the soluble form (Sβ-gal) with two different substrates (ONPG and PNPG) and at different aging times. Data were analyzed with a Michaellis-Menten model. With the substrate ONPG, we found a Vmax value higher for Eβ-gal than for Sβ-gal and this difference increased at longer aging times. On the contrary, with PNPG, Vmax was not affected neither by the enzyme condition (Sβ-gal and Eβ-gal) nor by the aging time. The porous structure of the silicate matrix was also studied as a function of aging time of the gels. Thus, fluorescence correlation spectroscopy (FCS) was applied through the analysis of diffusional properties of fluorescent probes of different sizes, entrapped in the gel matrix. We observed an anomalous diffusion in some conditions, suggesting a self-similar pore structure, and aging time dependent changes in diffusion coefficients. Taken together our result supports the hypothesis that there is a relationship between the catalytic mechanism proposed for β-gal and water availability in the silicate gel since the ratio between bulk and structured water depends on the pore size. Fil: Burgos, Martha Ines. 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: Carrer, Dolores Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; 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 XLII Reunión Anual de la Sociedad Argentina de Biofísica Villa Carlos Paz Argentina Sociedad Argentina de Biofísica |
| description |
Protein encapsulation in solid matrixes is of interest for biotechnological purposes and it also serves as a model of molecular crowding. We have successfully entrapped the enzyme β-galactosidase (β-gal) in silicate gels via a sol-gel reaction. The catalytic activity of encapsulated β-gal (Eβ-gal) was compared with the activity of the soluble form (Sβ-gal) with two different substrates (ONPG and PNPG) and at different aging times. Data were analyzed with a Michaellis-Menten model. With the substrate ONPG, we found a Vmax value higher for Eβ-gal than for Sβ-gal and this difference increased at longer aging times. On the contrary, with PNPG, Vmax was not affected neither by the enzyme condition (Sβ-gal and Eβ-gal) nor by the aging time. The porous structure of the silicate matrix was also studied as a function of aging time of the gels. Thus, fluorescence correlation spectroscopy (FCS) was applied through the analysis of diffusional properties of fluorescent probes of different sizes, entrapped in the gel matrix. We observed an anomalous diffusion in some conditions, suggesting a self-similar pore structure, and aging time dependent changes in diffusion coefficients. Taken together our result supports the hypothesis that there is a relationship between the catalytic mechanism proposed for β-gal and water availability in the silicate gel since the ratio between bulk and structured water depends on the pore size. |
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2013 |
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2013 |
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http://hdl.handle.net/11336/237833 Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging time; XLII Reunión Anual de la Sociedad Argentina de Biofísica; Villa Carlos Paz; Argentina; 2013; 59-59 978-987-27591-2-4 CONICET Digital CONICET |
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Catalytic activity of beta–galactosidase entrapped in a silicate matrix: Effect of the substrate type, porous structure and aging time; XLII Reunión Anual de la Sociedad Argentina de Biofísica; Villa Carlos Paz; Argentina; 2013; 59-59 978-987-27591-2-4 CONICET Digital CONICET |
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eng |
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Sociedad Argentina de Biofísica |
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