Extracción de solasodina de la especie Solanum marginatun usando β-Glucosidasa inmovilizada sobre nanopartículas magnéticas de CoFe2O4

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dc.contributor.advisorReyes Cuellar, Julia Constanzaspa
dc.contributor.authorGarcía Colmenares, José Mauricio
dc.date.accessioned2019-05-10T21:24:54Z
dc.date.available2019-05-10T21:24:54Z
dc.date.issued2017
dc.description1 recurso en línea (89 páginas) : ilustraciones color, figuras, tablas, imágenes.spa
dc.description.abstractA nivel industrial, el uso de enzimas inmovilizadas simplifica el proceso y disminuye el costo económico de la producción. Se investigó el reusó de la enzima β-glucosidasa inmovilizada sobre nanopartículas magnéticas de ferrita de cobalto CoFe2O4 activadas con glutaraldehído, para aplicarla en la hidrólisis de extractos de solasonina, provenientes de frutos semimaduros de la especie Solanum marginatum. El trabajo se dividió en cinco capítulos. En el capítulo I, se aborda el soporte teórico del trabajo de investigación. En el capítulo II, se presenta la síntesis y caracterización de las nanopartículas magnéticas de ferrita de cobalto (CoFe2O4). Se sintetizaron tres relaciones molares de nanopartículas. Se evaluó el rendimiento de síntesis y la atracción magnética, obteniendo mejores resultados con la relación Co40Fe60. Con estas nanopartículas, en el capítulo II, se realizó el proceso de inmovilización usando como modelo la enzima bromelina. La inmovilización se efectuó covalentemente mediante puentes de glutaraldehído y nanopartículas recubiertas con quitosano. Los resultados de este capítulo se condensan en un artículo titulado: Cinética de bromelina inmovilizada sobre nanopartículas magnéticas de cobalto-hierro (CoFe2O4), enviado a la revista South African Journal of Science, el cual se encuentra en evaluación. En el capítulo III, se realizó la aplicación del sistema de inmovilización y caracterización de la enzima β-glucosidasa sobre nanopartículas magnéticas de CoFe2O4 activadas con glutaraldehído. Se evaluaron cuatro tipos de inmovilización. La β-glucosidasa tipo A presentó la mayor eficiencia de inmovilización con 84,6 ± 2,1%. La actividad catalítica de la enzima libre e inmovilizada fue determinada en 93 ±2,6 y 82 ± 2,8% respectivamente. La inmovilización tipo D presentó el porcentaje de inmovilización más bajos con 25 ±2,4%. El sistema fue optimizado bajo las siguientes condiciones: 200 mg de NPM, glutaraldehído 1M, β-glucosidasa 5,7E-5M, tiempo de inmovilización 16 horas a 60 °C. La β-glucosidasa inmovilizada puede ser reutilizada durante diez ciclos manteniendo el 52 ± 2,5% de su actividad. La caracterización de las nanopartículas CoFe2O4 (Co40Fe60) y de la β-glucosidasa se evaluó por: microscopía electrónica de transmisión (TEM-STEM), difracción de rayos X (DRX) espectroscopias FT-IR y UV-vis, voltametría cíclica (VC), voltametría de onda cuadrada (VOC) y magnetometría de muestra vibrante (VSM). En el capítulo IV, se presenta la aplicación de la β-glucosidasa inmovilizada, en la hidrólisis de solasonina para obtención de solasodina a partir de frutos semimaduros de Solanum marginatum, logrando obtener un rendimiento de 1,8-2,5% y pureza de 89,2-91,3 %. Siendo mayor cando se realiza la extracción vía hidrólisis ácida. Sin embargo, esta diferencia es superada por la β-glucosidasa inmovilizada al lograr ser reutilizada durante diez ciclos y es amigable con el entorno natural. Como aporte, se realizó la caracterización por técnicas voltamperométricas aún no reportadas en la literatura, de las NPM-CoFe2O4 y de la β-glucosidasa inmovilizada. Se modificó el proceso de extracción de solasonina, fundamentado en la formación de sales de acetato de solasodina, haciéndolo amigable con el medio ambiente.spa
dc.description.abstractIndustrial use of immobilized enzyme simplifies the process and improves the economics and process efficiencies. The reuse of immobilized β-glucosidase enzyme on glutaraldehyde-activated cobalt ferrite magnetic nanoparticles was investigated for the hydrolysis of solasonin extracts from semi-mature fruits of Solanum marginatum. The work was divided into five chapters. In Chapter I, the theoretical support of the research work is addressed. In Chapter II, the synthesis and characterization of the cobalt ferrite magnetic nanoparticles (CoFe2O4) is presented. Three molar ratios of nanoparticles were synthesized. The performance of synthesis and magnetic attraction were evaluated, obtaining better results with the relation Co40Fe60. With these nanoparticles, in chapter II, the immobilization process was performed using the enzyme bromelain as a model. The immobilization was performed covalently by glutaraldehyde bridges and chitosan-coat. The results of this chapter are condensed in an article titled: Kinetics of immobilized bromelain on magnetic nanoparticles of cobalt-iron (CoFe2O4), sent to the South African Journal of Science, which is being evaluated. In Chapter III, the immobilization and characterization of β-glucosidase enzyme was applied to glutaraldehyde activated CoFe2O4 magnetic nanoparticles. Four types of immobilization were evaluated. Type A β-glucosidase presented the highest immobilization efficiency with 84.6 ± 2.1%. The catalytic activity of the free and immobilized enzyme was determined in 93 ± 2.6 and 82 ± 2.8% respectively. Type D immobilization showed the lowest immobilization percentage with 25 ± 2.4%. The system was optimized under the following conditions: 200 mg of MNP, 1M glutaraldehyde, 5E-5M β-glucosidase, immobilization time 16 hours at 60 ° C. Immobilized β-glucosidase can be reused for ten cycles while maintaining 52 ± 2.5% of its activity.The characterization of the nanoparticles CoFe2O4 (Co40Fe60) and β-glucosidase was evaluated by: transmission electron microscopy (TEM-STEM), X-ray diffraction (XRD) FT-IR and UV-vis spectroscopies, cyclic voltammetry (CV) , square wave voltammetry (SWV) and vibration sample magnetometry (VSM). In Chapter IV, the application of the immobilized β-glucosidase, in the hydrolysis of solasonin to obtain solasodine from semi-mature fruits of Solanum marginatum, is presented, obtaining a yield of 1.8-2.5% and purity of 89.2-91.3%. Being greater when the extraction is carried out via acid hydrolysis. However, this difference is overcome by the immobilized β-glucosidase to be reused for ten cycles and is friendly to the natural environment. As a contribution, characterization was performed by voltamperometric techniques not yet reported in the literature, of NPM-CoFe2O4 and immobilized β-glucosidase. The extraction process of solasonin, based on the formation of salts of acetate of solasodine, was modified, making it friendly with the environment.eng
dc.description.degreelevelMaestríaspa
dc.description.degreenameMagíster en Químicaspa
dc.description.notesBibliografía: páginas 83-87.spa
dc.format.mimetypeapplication/pdfspa
dc.identifier.citationGarcía Colmenares, J. M. (2017). Extracción de Solasodina de la especie Solanum marginatun usando β-Glucosidasa inmovilizada sobre nanopartículas magnéticas de CoFe2O4. (Tesis de Maestría). Universidad Pedagógica y Tecnológica de Colombia, Tunja. http://repositorio.uptc.edu.co/handle/001/2582spa
dc.identifier.urihttp://repositorio.uptc.edu.co/handle/001/2582
dc.language.isospaspa
dc.publisherUniversidad Pedagógica y Tecnológica de Colombiaspa
dc.publisher.facultyFacultad de Ciencias. Escuela de Posgrados. Maestría en Químicaspa
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dc.rightsCopyright (c) 2017 Universidad Pedagógica y Tecnológica de Colombiaspa
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dc.rights.creativecommonsAtribución-NoComercial 4.0 Internacional (CC BY-NC 4.0)spa
dc.rights.urihttps://creativecommons.org/licenses/by-nc/4.0/spa
dc.subject.armarcEnzimas inmovilizadas
dc.subject.armarcBiorreactores
dc.subject.armarcEnzimas
dc.subject.armarcEnzimas Inmovilizadas - Biotecnología
dc.subject.armarcMaestría en Química - Tesis y disertaciones académicas
dc.titleExtracción de solasodina de la especie Solanum marginatun usando β-Glucosidasa inmovilizada sobre nanopartículas magnéticas de CoFe2O4spa
dc.typeTrabajo de grado - Maestríaspa
dc.type.coarhttp://purl.org/coar/resource_type/c_bdccspa
dc.type.coarversionhttp://purl.org/coar/version/c_970fb48d4fbd8a85spa
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dc.type.redcolhttps://purl.org/redcol/resource_type/TMspa
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