Biotecnología

 

Caracterización del componente prebiótico de Aspergillus niger estimulante de bacterias celulóticas de origen ruminal

 

Characterization of the prebiotic component of Aspergillus niger that stimulates the celullolytic bacteria of ruminal origin

 

A. Quintero–Lira¹, R. Hernández–Díaz¹, D. J. Pimentel–González¹, M. E. Rodríguez–Huezo² y R. G. Campos–Montiel¹*

 

¹ ICAP, Universidad Autónoma del Estado de Hidalgo, Av. Universidad Km. 1, Rancho Universitario, 43600, Tulancingo, Hgo., México. * Autor para la correspondencia. E–mail: rcampos@uaeh.reduaeh.mx Tel. (771) 71 7 20 00, ext. 4641; Fax: (771) 71 7 20 000

² Depto. Ingeniería Química y Bioquímica, Tecnológico de Estudios Superiores de Ecatepec, Av. Tecnológico s/n esq. Av. Central, Col. Valle de Anáhuac, 55210, Ecatepec, Edo. Méx., México.

 

Recibido 12 de Noviembre 2008
Aceptado 15 de Enero 2009

 

Resumen

Extractos crudos de Aspergillus niger (An) se han utilizado como aditivos para la nutrición animal demostrando su efecto al incrementar la digestibilidad de forrajes en ensayos in vitro. En esta investigación se caracterizó el componente prebiótico de An que estimula el crecimiento de un consorcio de bacterias celulolíticas de origen ruminal (CBCOR). Se utilizaron fracciones ultrafiltradas menores de 30 kDa. Las fracciones ultrafiltradas fueron calentadas a diferentes temperaturas (60, 90 y 120 °C), se estimó su peso molecular mediante cromatografía de exclusión molecular y se fraccionaron con cromatografía de intercambio iónico. A las fracciones activas se les realizó análisis de proteína, carbohidratos y espectrofotometría de infrarrojo. Los resultados mostraron que todas las fracciones ultrafiltradas y tratadas térmicamente incrementaron significativamente (P<0.05) la producción de proteína, concentración de acetato y la actividad carboximetilcelulasas en CBCOR. La cromatografía de exclusión molecular estimó un peso molecular de 6.1 kDa de la fracción activa, mientras las cromatografías de intercambio aniónico y catiónico mostraron una sola fracción activa. El análisis bioquímico y espectro infrarrojo sugieren la naturaleza peptídica del componente prebiótico de An que estimula el CBCOR.

Palabras clave: prebióticos, bacterias anaerobias celulolíticas, Aspergillus niger.

 

Abstract

It has been demonstrated that crude extracts of Aspergillus niger (An) used as additives for animal nutrition have enhanced the digestibility of forages on in vitro assays. In this work the prebiotic component of An that stimulates the growth of a cellulolitic bacterial consortium from ruminal origin (CBCRO) was characterized. Ultrafiltrated fractions of less than 30 kDa, heat treated at 60, 90 and 120 °C were used. The fractions were obtained by ion–exchange chromatography and their average molecular weight was determined by molecular exclusion chromatography. Protein and carbohydrate contents and the infrared spectra were determined on the active fractions. Results showed that all the heat treated ultrafiltrated fractions significantly increased (P<0.05) carboxymethylcellulases' activities, and protein and acetate productions of the CBCRO. Molecular exclusion chromatography established that the active fraction had a molecular mass of 6.1 kDa, whereas only one active fraction was detected through ion exchange chromatography. Biochemical analysis and infrared spectra data suggest that the prebiotic component of An has a peptidic nature.

Keywords: prebiotics, cellulolytic anaerobic bacteria, Aspergillus niger.

 

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