Actividad celulolítica de hongos filamentosos aislados del Parque Nacional de Foz de Iguaçu – Brasil en sustratos agrícolas mediante la planificación experimental
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2020-03-04
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Rodríguez Andachi, Anabel Alexandra
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Las celulasas son un conjunto de enzima hidrolíticas que descomponen la celulosa, mediante la actividad sinérgica de tres diferentes componentes, las endoglucanasa, exoglucanasa y la β -glucosidasa. A lo largo de los años estudios de celulasas han demostrado ser potencial biotecnológico en diferentes áreas, aplicada principalmente por ser biodegradables e inofensivas con el medio ambiente. Su producción es bastante estudiada en residuos agrícolas o agroindustriales. La creciente industrialización de estas áreas, ha llevado a la producción de altas cantidades de residuos que no son tratados adecuadamente. Siendo importante el aislamiento de microorganismos con potenciales celulolíticos. Esa posibilidad es brindada por la exploración del Parque Nacional Foz de Iguaçu que, por su biodiversidad vegetal, materia orgánica y minerales en el suelo, pudieron haber seleccionado microorganismos con potencial, diferentes a los ya descritos. Así, este trabajo tiene como objetivo evaluar el potencial de degradación de celulosa por fungos del PNI en sustratos agrícolas mediante planificación experimental. Y estudio previo de 30 fungos presentaron degradación de celulosa, determinado por la actividad, cuando cultivados en CMC sólido. Siendo así, fueron cultivados en CMC líquido y analizado bajo el protocolo Kit CellG5 cellulase assay Megazyme. El fungo Fusarium sp. CCLM CX presentó la mayor actividad enzimática (1,729 U CellG5/mL), el cual fue seleccionado para la evaluación frente a los sustratos naturales (salvado de trigo, salvado de avena, mazorca de maíz y aserrín). Para este estudio fue ejecutado una planificación experimental tipo fraccional 25-1. En sustratos naturales, la actividad enzimática del fungo CX fue menor que en CMC, ya que CMC está lista para el consumo metabólico, mientras que los sustratos tienen una composición compleja para el acceso al carbono. Por otro lado, los sustratos que tuvieron efecto significativo en el análisis estadístico fueron, el salvado de trigo y mazorca de maíz. Por los tanto, se deben realizar nuevos experimentos, con el fungo CX, conociendo el sustrato principal para la optimización enzimática. Las propiedades químicas de estos sustratos pueden ser aplicados principalmente como fuentes de producción de celulasas y aplicación en dieta animal o como biofertilizante, en regiones como Paraná.
Cellulases are a set of hydrolytic enzymes that break down cellulose, through the synergistic activity of three different components, endoglycanase, exoglycanase and β-glucosidase. Over the year’s studies of cellulases have proven to be biotechnological potential in different areas, mainly applied because they are biodegradable and harmless to the environment. Its production is quite studied in agricultural or agro-industrial waste. The increasing industrialization of these areas has led to the production of high amounts of waste that are not treated properly. The isolation of microorganisms with cellulolytic potentials is important. This possibility is provided by the exploration of the Foz de Iguaçu National Park which, due to its plant biodiversity, organic matter and minerals in the soil, may have selected microorganisms with potential, different from those already described. Thus, this work aims to evaluate the potential for cellulose degradation by fungi of the PNI in agricultural substrates through experimental planning. And a previous study of 30 fungi showed cellulose degradation, determined by activity, when grown in solid CMC. Thus, they were cultured in liquid CMC and analyzed under the Kit CellG5 cellulase assay Megazyme protocol. The fungus Fusarium sp. CCLM CX had the highest enzymatic activity (1,729 U CellG5 / mL), which was selected for evaluation against natural substrates (wheat bran, oat bran, corn cob and sawdust). For this study, a fractional 2 5-1 experimental planning was executed. In natural substrates, the enzymatic activity of the CX fungus was lower than in CMC, since CMC is ready for metabolic consumption, while the substrates have a complex composition for access to carbon. On the other hand, the substrates that had a significant effect on the statistical analysis were wheat bran and corn cob. Therefore, new experiments must be carried out, with the CX fungus, knowing the main substrate for enzymatic optimization. The chemical properties of these substrates can be applied mainly as sources of cellulase production and application in animal diet or as biofertilizer, in regions such as Paraná.
Cellulases are a set of hydrolytic enzymes that break down cellulose, through the synergistic activity of three different components, endoglycanase, exoglycanase and β-glucosidase. Over the year’s studies of cellulases have proven to be biotechnological potential in different areas, mainly applied because they are biodegradable and harmless to the environment. Its production is quite studied in agricultural or agro-industrial waste. The increasing industrialization of these areas has led to the production of high amounts of waste that are not treated properly. The isolation of microorganisms with cellulolytic potentials is important. This possibility is provided by the exploration of the Foz de Iguaçu National Park which, due to its plant biodiversity, organic matter and minerals in the soil, may have selected microorganisms with potential, different from those already described. Thus, this work aims to evaluate the potential for cellulose degradation by fungi of the PNI in agricultural substrates through experimental planning. And a previous study of 30 fungi showed cellulose degradation, determined by activity, when grown in solid CMC. Thus, they were cultured in liquid CMC and analyzed under the Kit CellG5 cellulase assay Megazyme protocol. The fungus Fusarium sp. CCLM CX had the highest enzymatic activity (1,729 U CellG5 / mL), which was selected for evaluation against natural substrates (wheat bran, oat bran, corn cob and sawdust). For this study, a fractional 2 5-1 experimental planning was executed. In natural substrates, the enzymatic activity of the CX fungus was lower than in CMC, since CMC is ready for metabolic consumption, while the substrates have a complex composition for access to carbon. On the other hand, the substrates that had a significant effect on the statistical analysis were wheat bran and corn cob. Therefore, new experiments must be carried out, with the CX fungus, knowing the main substrate for enzymatic optimization. The chemical properties of these substrates can be applied mainly as sources of cellulase production and application in animal diet or as biofertilizer, in regions such as Paraná.
Descrição
Trabajo de Conclusión de Curso II presentado al
Instituto Latinoamericano de Ciencias de la Vida y de la
Naturaleza de la Universidad Federal de la Integración
Latinoamericana, como requisito parcial a la obtención
del título de grado en Biotecnología.
Orientadora: Profa. Dra. Rafaella Costa Bonugli Santos
Co-orientador: Prof. Dr. Michel Passarini
Palavras-chave
Celulasas, Hongos productores de celulasas, Degradación de celulosa
Citação
RODRÍGUEZ ANDACHI, Anabel Alexandra. Actividad celulolítica de hongos filamentosos aislados del Parque Nacional de Foz de Iguaçu – Brasil en sustratos agrícolas mediante la planificación experimental. 2019. 54 p. Trabajo de Conclusión de Carrera (Bacharel em Biotecnología) – Universidad Federal de la integración Latinoamericana, Foz de Iguaçu, 2019.