Producción y caracterización de las enzimas lignocelulolíticas del hongo de pudrición blanca de la madera Bjerkandera adusta en cocultivo con el hongo de pudrición parda Gloeophyllum trabeum
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Date
2024
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Universidad de Concepción
Abstract
El presente estudio investigó la interacción entre dos hongos pudridores de la madera, Bjerkandera adusta, de pudrición blanca y Gloeophyllum trabeum, de pudrición parda, en la producción de enzimas lignocelulolíticas, hidrolíticas y oxidativas, mediante cocultivo bajo condiciones de fermentación líquida. Para investigar la compatibilidad entre las especies, inicialmente se evaluó el crecimiento de las especies en medio sólido, donde fue posible observar un crecimiento micelial entremezclado de ambos hongos, sin producción de pigmento en la interfaz de los micelios, indicando una compatibilidad parcial, además se obtuvo que el cocultivo
no inhibió el crecimiento micelial de ambos hongos, y en algunos casos, promovió un ligero aumento del crecimiento en comparación con el monocultivo. La caracterización de los cocteles enzimáticos concentrados de B. adusta en cocultivo en medio líquido, proporcionó una visión detallada de las interacciones y la influencia de G. trabeum en su actividad enzimática. En términos de producción enzimática, se registró un aumento significativo en la actividad de las enzimas oxidativas en el cocultivo, destacando un incremento de 2-3 veces en la producción
de peroxidasas, respecto al monocultivo de B. adusta. Estos resultados sugieren el efecto estimulante de G. trabeum (que no produce peroxidasas) sobre B. adusta en cocultivo. Por el contrario, en el caso de las enzimas celulolíticas y hemicelulolíticas se observó una significativa inhibición por efecto del cocultivo, respecto al monocultivo de G. trabeum. Las técnicas de electroforesis nativa y zimograma confirmaron la presencia y actividad de peroxidasas sólo en B. adusta y el cocultivo, y su ausencia en G. trabeum, y la presencia de xilanasas sólo en G. trabeum. La purificación de los extractos por cromatografía de intercambio iónico reveló la
separación de diferentes fracciones proteicas, algunas de las cuales presentaron actividad de endoglucanasa (en todos los cultivos) y peroxidasas (monocultivo de B. adusta y cocultivo). Estos resultados contribuyen a comprender las interacciones fúngicas, en la producción de enzimas lignocelulolíticas, destacando el potencial del cocultivo para aumentar la producción de enzimas oxidativas en B. adusta con diversos fines biotecnológicos
The present study investigated the interaction between two wood-rotting fungi, Bjerkandera adusta, white rot, and Gloeophyllum trabeum, brown rot, in the production of lignocellulolytic (hydrolytic and oxidative) enzymes by coculture under liquid fermentation conditions. To study the compatibility between the species, their growth was first evaluated in solid medium, where a mixed mycelial growth of both fungi was observed, without pigment production at the interface of the mycelia, indicating a partial compatibility. In addition, it was found obtained that the coculture did not inhibit the mycelial growth of either fungus, and in some cases, promoted a slight increase in growth compared to the monoculture. Characterization of the concentrated enzyme cocktails of B. adusta in coculture in liquid medium provided detailed insight into the interactions and influence of G. trabeum on their enzymatic activity. In terms of enzyme production, a significant increase in the activity of oxidative enzymes was observed in the coculture, highlighting a 2-3-fold increase in the production of peroxidases in the coculture compared to B. adusta monoculture. These results suggest a stimulatory effect of G. trabeum (which does not produce peroxidases) on B. adusta in coculture. On the contrary, in the case of cellulolytic and hemicellulolytic enzymes, a significant inhibition was observed due to the effect of the coculture, with respect to the monoculture of G. trabeum. Native electrophoresis and zymogram techniques confirmed the presence and activity of peroxidases only in B. adusta and coculture, and their absence in G. trabeum, and the presence of xylanases only in G. trabeum. Purification of the extracts by ion exchange chromatography revealed the separation of different protein fractions, some of which showed endoglucanase activity (in all cultures) and peroxidases (B. adusta and coculture). These results contribute to the understanding of fungal interactions in the production of lignocellulolytic enzymes and highlight the potential of coculture to increase the production of oxidative enzymes in B. adusta for various biotechnological purposes
The present study investigated the interaction between two wood-rotting fungi, Bjerkandera adusta, white rot, and Gloeophyllum trabeum, brown rot, in the production of lignocellulolytic (hydrolytic and oxidative) enzymes by coculture under liquid fermentation conditions. To study the compatibility between the species, their growth was first evaluated in solid medium, where a mixed mycelial growth of both fungi was observed, without pigment production at the interface of the mycelia, indicating a partial compatibility. In addition, it was found obtained that the coculture did not inhibit the mycelial growth of either fungus, and in some cases, promoted a slight increase in growth compared to the monoculture. Characterization of the concentrated enzyme cocktails of B. adusta in coculture in liquid medium provided detailed insight into the interactions and influence of G. trabeum on their enzymatic activity. In terms of enzyme production, a significant increase in the activity of oxidative enzymes was observed in the coculture, highlighting a 2-3-fold increase in the production of peroxidases in the coculture compared to B. adusta monoculture. These results suggest a stimulatory effect of G. trabeum (which does not produce peroxidases) on B. adusta in coculture. On the contrary, in the case of cellulolytic and hemicellulolytic enzymes, a significant inhibition was observed due to the effect of the coculture, with respect to the monoculture of G. trabeum. Native electrophoresis and zymogram techniques confirmed the presence and activity of peroxidases only in B. adusta and coculture, and their absence in G. trabeum, and the presence of xylanases only in G. trabeum. Purification of the extracts by ion exchange chromatography revealed the separation of different protein fractions, some of which showed endoglucanase activity (in all cultures) and peroxidases (B. adusta and coculture). These results contribute to the understanding of fungal interactions in the production of lignocellulolytic enzymes and highlight the potential of coculture to increase the production of oxidative enzymes in B. adusta for various biotechnological purposes
Description
Tesis para optar al grado de Ingeniero en Biotecnología Vegetal