Optimización de la producción de metabolitos secundarios tipo flavonoides en cultivos en suspensión de células de Thevetia peruviana

dc.contributorArias Zabala, Mario Evelio
dc.contributorUniversidad Nacional de Colombia - Sede Medellín
dc.contributorBiotecnología Industrial
dc.creatorLeón Durán, Mateo David
dc.date.accessioned2020-08-25T21:09:07Z
dc.date.available2020-08-25T21:09:07Z
dc.date.created2020-08-25T21:09:07Z
dc.date.issued2020-08-21
dc.identifierLeón Durán, Mateo David (2020) Optimización de la producción de metabolitos secundarios tipo flavonoides en cultivos en suspensión de células de Thevetia peruviana. Tesis de Maestría. Universidad Nacional de Colombia - sede Medellín
dc.identifierhttps://repositorio.unal.edu.co/handle/unal/78225
dc.description.abstractPlants are the main source for obtaining secondary metabolites that are used in the pharmaceutical, cosmetic and food industries. In nature, the performance of bioactive compounds is affected by biotic and abiotic factors, an alternative to overcome this adversity is in vitro plant cultures and particularly plant cell culture that has multiple advantages, highlighting the possibility of controlling variables to increase the content. of compounds of interest. The objective of this research was to determine and optimize the effect of some elicitors on the production of secondary flavonoid metabolites in suspension cell cultures of Thevetia peruviana on a shake flask scale. The experimental part was performed from cell cultures in suspension cells of T. peruviana maintained by the Bioconversion Laboratory of the Universidad Nacional de Colombia, sede Medellin. Firstly, methyl jasmonate (MeJa) 3 µM and salicylic acid (SA) 300 µM were added to two los of cell cultures, that circumstantially they differed in their time suspension state lot 1 (9 months) and lot 2 (3 months). Secondly, certain were evaluated, in order to be optimized; the operational parameters cosidered were: day of addition, concentration and hours of harvest. Finally, the combined effect of MeJa (0.3 µM) and SA (100 µM) was determined in different proportions, the day of elicitation and the hours of harvest were constant. Flavonoid content was quantified by UV-vis spectrophotometry usinfg the AlCl3 complexation method and was evaluated at extracellular and intracellular level. In the first part, at intracellular level, SA generated 14.6 % and 9.56 % more flavonoid content than MeJa in lots 1 and 2, respectively. In the second part, 4.14 mg EQ/g DW were generated with MeJa 0.3 µM, elicitation day 5 and harvest at 90 h; 3.75 mg EQ/g DW were generated with 100 µM SA, elicitation day 0 and harvests at 96 h. In the third part, 4.62 mg EQ / g DW were generated with the combination of MeJa (0.3 µM) – SA (100 µM) in a ratio of 20-80, elicitation on day 0 and harvest at 96 h. Eliciting cell cultures of Thevetia peruviana with MeJa and SA under optimal conditions of concentration, day of addition and hours of harvest increased the content of flavonoid compounds. The results obtained could serve as a basis for the development of investigations at the bioreactor scale.
dc.description.abstractLas plantas son la principal fuente para la obtención de metabolitos secundarios que se usan en la industria farmacéutica, cosmética y alimentaria. En la naturaleza el rendimiento de compuestos bioactivos se ve afectado por factores bióticos y abióticos, una alternativa para superar esta adversidad son los cultivos vegetales in vitro y particularmente el cultivo de células en suspensión que presenta múltiples ventajas destacándose la posibilidad de controlar variables para aumentar el contenido de compuestos de interés. El objetivo del presente trabajo de investigación fue determinar y optimizar el efecto de algunos elicitores sobre la producción de metabolitos secundarios tipo flavonoides en cultivos de células en suspensión de Thevetia peruviana a escala de matraz agitado. La parte experimental se realizó a partir de cultivos de células en células en suspensión de T. peruviana que mantenía el laboratorio de bioconversiones de la Universidad Nacional de Colombia sede Medellín. Primeramente, metil jasmonato (MeJa) 3 μM y ácido salicílico (AS) 300 μM se adicionaron a dos lotes de cultivos de células; que circunstancialmente diferían en su tiempo en estado de suspensión lote 1 (9 meses) y lote 2 (3 meses). Segundamente, determinadas condiciones fueron evaluadas, con el fin de ser optimizadas; los parámetros operacionales que se consideraron fueron: día de adición, concentración y las horas de cosecha. Por último, el efecto combinado de MeJa (0,3 μM) – AS (100 μM) se determinó en diferentes proporciones, el día de elicitación y las horas de cosecha fueron constantes. El contenido de flavonoides se cuantificó por espectrofotometría UV – vis por el método de complejación de AlCl3 y se evaluaron a nivel extracelular e intracelular. En la primera parte, a nivel intracelular AS generaron 14,6 % y 9,56 % más contenido de flavonoides que MeJa en lotes 1 y 2, respectivamente. En la segunda parte, 4,14 mg EQ/g MS fueron generados con MeJa 0,3 μM, elicitación día 5 y cosecha a 90 h; por otro lado, 3,75 mg EQ/g MS fueron generados con AS 100 μM, elicitación día 0 y cosecha a 96 h. En la tercera parte, 4,62 mg EQ/g MS fueron generados con la combinación de MeJa (0,3 μM) – AS (100 μM) en proporción 20 – 80, elicitación el día 0 y cosecha a las 96 h. Elicitar cultivos celulares de Thevetia peruviana con MeJa y AS en condiciones óptimas de concentración, día de adición y horas de cosecha incrementaron el contenido de compuestos flavonoides. Los resultados obtenidos podrían servir como base para el desarrollo de investigaciones a escala de biorreactor.
dc.languagespa
dc.publisherMedellín - Ciencias - Maestría en Ciencias - Biotecnología
dc.publisherEscuela de biociencias
dc.publisherUniversidad Nacional de Colombia - Sede Medellín
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dc.rightsAtribución-NoComercial-SinDerivadas 4.0 Internacional
dc.rightsAcceso abierto
dc.rightshttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rightsinfo:eu-repo/semantics/openAccess
dc.rightsDerechos reservados - Universidad Nacional de Colombia
dc.titleOptimización de la producción de metabolitos secundarios tipo flavonoides en cultivos en suspensión de células de Thevetia peruviana
dc.typeOtro


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