Identificación de β-galactosidasa, β-fructofuranosidasa y glicosil transferasa de Cellulomonas flavigena al crecer en diferentes fuentes de carbono
Identification of β-galactosidase, β-fructofuranosidase and glycosiltransferase enzymes from Cellulomonas flavigena when grown in several carbon sources
E. Solís-Badillo1, L. Mayorga-Reyes2, A. Azaola-Espinosa2*, A. Gutiérrez-Nava2
1 Maestría en Ciencias Agropecuarias. Universidad Autónoma Metropolitana-Xochimilco. Calzada del Hueso 1100, Col. Villa Quietud, Del. Coyoacán. C. P. 04960 México, D. F.
2 Dpto. de Sistemas Biológicos. Universidad Autónoma Metropolitana-Xochimilco. Calzada del Hueso 1100, Col. Villa Quietud, Del. Coyoacán. C. P. 04960 México, D. F. * Corresponding author. E-mail: azaola@correo.xoc.uam.mx Tel. 55-54-83-73-77
]]>Received September 4, 2013.
Accepted December 31, 2013.
Resumen
Cellulomonas produce una amplia variedad de glucanasas, entre estas las celulasas y hemicelulasas son las más sobresalientes por sus aplicaciones industriales. Sin embargo, ha mostrado la capacidad de crecer en sustratos diferentes a los derivados de residuos lignocelulósicos. C. flavigena creció lactosa, sacarosa y raftilosa. En lactosa y sacarosa la producción de células fue de aproximadamente 2 mg/mL y en raftilosa solo alcanzó 1 mg/mL. Se determinó la actividad hidrolítica en los extractos intracelulares y extracelulares correspondientes a las actividades de β-galactosidasa y β-fructofuranosidasa. La mayor actividad hidrolítica se encontró en los extractos intracelulares cuando se utilizó raftilosa como sustrato y disminuyó significativamente al utilizar lactosa. Los perfiles de proteínas de los distintos extractos intracelulares presentan gran cantidad de bandas en los distintos sustratos utilizados como fuente de carbono. Probablemente en cada de las fuentes estudiadas, la síntesis de enzimas intracelulares no esté regulada y se exprese todo el potencial de esta bacteria. Además se identificó la presencia del gen de la β-galactosidasa.
Palabras clave: Cellulomonas flavigena, β-galactosidasa, β-fructofuranosidasa, bifuncionalidad.
Abstract
The genus Cellulomonas is a cellulose degrading bacteria which produce a wide variety of glucanases, mainly cellulases and hemicellulases with industrial applications. C. flavigena has the ability to grow on carbon sources that are not derived from lignocellulosic wastes such as lactose, sucrose and raftilose. In this study, when C. flavigena was cultured on lactose and sucrose, the cell production was approximately 2 mg/mL, and when was cultured on raftilose, it reached 1 mg/mL. The hydrolytic activity of β-galactosidase and β-fructofuranosidase enzymes were tested in the intra and extra cellular fractions where the highest activity was found in the intracellular fraction using raftilose as substrate, nevertheless using lactose as substrate the activity was significantly low. The protein profiles of the intracellular extracts presented great quantity of bands in the different substrates used as carbon sources of carbon. Probably the synthesis of intracellular enzymes is not regulated and the cell expresses its whole potential. In addition the presence of the gene β-galactosidase was identified.
]]> Keywords: Cellulomonas flavigena, β-galactosidase, β-fructofuranosidase, bifunctionality.
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Agradecimientos
ESB agradece al Consejo Nacional de Ciencia y Tecnología (CONACyT) por el apoyo otorgado durante la realización de los estudios de Maestría (367516).
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