pulpa de café por cepas seleccionadas de pulpa de café por cepas seleccionadas de pulpa de café por cepas seleccionadas de
% Muestra Nitrógeno
I.4.6. Zimogramas de las actividades enzimáticas ensayadas
En la Figura I.13 se muestran los zimogramas correspondientes a las distintas actividades enzimáticas ensayadas.
En la mayor parte de los casos analizados, se detectaron varias bandas de actividad. Este hecho es relativamente frecuente en cepas de Streptomyces cultivadas tanto en cultivo sumergido como en SSF. Así, en los zimogramas de actividad xilanasa correspondientes al sobrenadante de cultivo de la cepa S. chattanoogensis en cultivo sumergido, también se obtuvieron distintos patrones de bandas de actividad con diferente movilidad electroforética a distintos días de incubación, que se atribuyeron a la producción de distintas isoenzimas (López-Fernández, 1995). Este mismo comportamiento ha sido también observado para las mananasas producidas por S. ipomoea (Montiel y col., 2002). En los zimogramas de actividad lacasa realizados con los extractos procedentes de la pulpa de café fermentada con la cepa S. UAH Nic-C (Figura I.13.D) se obtuvieron dos bandas de actividad, que podrían corresponder a dos isoformas. En estudios realizados en nuestro laboratorio, se puso de manifiesto que la actividad lacasa producida por S. cyaneus en SSF sobre paja de trigo, se debe a la existencia de tres isoenzimas (Berrocal y col., 2000). Este patrón multibandas también ha sido descrito para lacasas de hongos (Palmieri y col., 2000).
50
Figura I.13 Zimogramas correspondientes a las actividades enzimáticas xilanasa (A), mananasa
(B) y amilasa (C) producidas por S. UAH Nic-C (1), S. UAH 47 (2) y S. chattanoogensis (3) y
lacasa (D) producida por la cepa S. UAH Nic C.
Del análisis de los resultados obtenidos en el estudio de las distintas actividades enzimáticas, se podría deducir, que la liberación de azúcares del almidón de la yuca por acción de las amilasas producidas, sustenta el crecimiento inicial de las cepas sobre el residuo pulpa de café, y una vez producida la biomasa e inducidas las hemicelulasas y lacasas, ya se podrían degradar otros componentes de la pulpa como las hemicelulosas y los polifenoles, lo que contribuye en este último caso, tal como se ha comentado anteriormente a su destoxificación, a la vez que se produce, un enriquecimiento proteico de los mismos (Tabla I.7). Como ya se ha sugerido, será necesario realizar estudios posteriores para comprobar si en períodos más largos de incubación del que se llevó a cabo el ensayo de las actividades enzimáticas (3 días), se producen en los sustratos fermentados por las cepas S. UAH 47 y S. chattanoogensis actividades de tipo oxidativo como la lacasa y/o la peroxidasa, actividades que se han implicado en la degradación de polímeros fenólicos como la lignina (Rob, y col. 1997; Berrocal, y col., 2000), y que aunque en un período tan corto no se han detectado, podrían ser la causa de la disminución del contenido en compuestos fenólicos determinado por Py- GC/MS. A A A A 2 1 3 B B B B D D D D C C C C
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