Biotecnología

 

Xylanolytic system proteins desorption from Aspergillus flavipes FP-500 cultures with agrowastes

 

Desorción de proteínas del sistema xilanolítico de Aspergillus flavipes FP-500 producidas en cultivos con residuos agro-industriales

 

L.R. Torres-Barajas y G. Aguilar-Osorio*

 

Departamento de Alimentos y Biotecnología. Conjunto E, Facultad de Química. Universidad Nacional Autónoma de México (UNAM). (Ciudad Universitaria. 04510 Coyoacán, México D.F. México. * Autor para la correspondencia. E-mail: gao@unam.mx.

 

Recibido 22 de Febrero de 2013
Aceptado 8 de Mayo de 2013

 

Resumen

Las enzimas degradadoras de carbohidratos que los hongos del genero Aspergillus son capaces de producir, han cobrado gran interés debido, entre otras cosas, a su aplicación en los procesos de obtención de etanol a partir de materias primas de bajo costo, como los residuos agroindustriales. Buena parte de las proteínas del sistema xilanolítico secretadas durante el cultivo de Aspergillus flavipes FP-500 en olote de maíz son adsorbidas en el sólido residual, a diferencia de las que se producen en cultivos con salvado de trigo, donde prácticamente no se adsorben. La recuperación de proteínas con soluciones de lavado a partir de ambos residuos indica que el incremento del pH favorece la desorción de las proteínas. Así mismo, la adsorción de las proteínas del sistema xilanolítico es más selectiva en salvado de trigo que en olote de maíz y puede verse influenciada por más de un mecanismo, ya que además de la adsorción debida a la interacción aleatoria de las proteínas con la superficie de los residuos, estas también podrían establecer interacciones específicas con ciertas regiones expuestas de los polisacáridos debidos a la presencia de dominios de unión a carbohidrato en la secuencia de algunas de las proteínas involucradas.

Palabras clave: xilanasas, olote de maíz, proteína, desorción, Aspergillus.

 

Abstract

Carbohydrate degrading enzymes produced by fungus such as Aspergillus are interesting due to its application into process of ethanol production using cheap materials such as agro-wastes. Proteins of xylanolytic system which are secreted during Aspergillus flavipes FP-500 culture with corn cobs as a carbon source, are adsorbed in the solid matrix of cobs unlike wheat bran in which virtually are non-absorbed. Recovery of proteins with washing solutions from both agro-wastes and protein sequences of some of these proteins shows: that increase of pH improves protein desorption, adsorption of xylanolytic system proteins is pretty much selective in wheat bran than in corn cobs and the adsorption of that proteins could be influenced by random interaction between protein and polysaccharide surface as well as specific interactions of proteins with polysaccharides by carbohydrate binding domains of certain involved proteins.

Keywords: xylanases, corn cobs, protein, desorption, Aspergillus.

 

DESCARGAR ARTÍCULO EN FORMATO PDF

 

Referencias

Arora, N., Kumar, A., Mutyala, S., Suryanarayana, U. (2009). Comparative characterization of commercially important xylanase enzymes. Bioinformation 3, 446-453.         [ Links ]

Benett, J.W. (1998). Review article. Mycotechnology: The Role of fungi in fitotechnology. Journal of Biotechnology 66, 101-107.         [ Links ]

Black, G., Rixon, J., Clarke, J., Hazlewood, G., Ferreira, L., Bolam, D., Gilbert, H. (1997). Cellulose binding domains and linkersequences potentiate the activity of hemicellulases against complex substrates. Journal of Biotechnology 57, 59-69.         [ Links ]

Bolio, G., Valadez, A., Veleva, L., Andreeva, A. (2011). Cellulose whiskers from agro-industrial banana wastes: isolation and characterization. Revista Mexicana de Ingeniería Química 10, 291-299.         [ Links ]

Cantarel, B., Coutinho, P., Rancurel, C., Bernard, T., Lombard, V., Henrissat, B. (2009). The Carbohydrate-Active EnZyme database (CAZy): An expert resource forglycogenomics. Nucleic Acids Research 37, D233-D238. (Carbohydrate Active Enzymes database, disponible en http://www.cazy.org/).         [ Links ]

Demirbas, A. (2008). Heavy metal adsorption onto agro-based waste materials: A review. Journal of Hazardous Materials 157, 220-229.         [ Links ]

Douglas, J., Johnson, H., Granick, S. (1993). A simple kinetic model ofpolymeradsorption and desorption. Science 262, 2010-2012.         [ Links ]

Farinella, N.V., Matos, G.D., Arruda, M.A.Z. (2007). Grape bagasse as potential biosorbent of metals in effluent treatments. Bioresource Technology 98, 1940-1946.         [ Links ]

Feramisco, J., Burridge, K. (1980). A rapid purification of a-actinin, filamin and 130,000-dalton protein from smooth muscle. The Journal of Biological Chemistry 255, 1194-1199.         [ Links ]

Fernandes, A., Fontes, C., Gilbert, H., Hazlewood, G., Fernandes T., Ferreira, L. (1999). Homologuos xylanases from Clostridium thermocellum: evidence for bifunctional activity, synergism between xylanase catalytic modules and the presence of xylan-binding domains in enzyme complexes. Biochemistry Journal 342, 105-110.         [ Links ]

Flores, T., Gutiérrez, M., Revah, S., Favela, E. (2011). Estudio comparativo de oxigenasas producidas por Aspergillus niger ATCC 9642 en fermentacioín en estado soílido y sumergido. Revista Mexicana de Ingeniería Química 10, 189-207.         [ Links ]

Gonçalves, F., Gonçalves, A., Gonçalves, E., Azevedo, M., Pinto, B., Duque, P., Sobral, L., Souza, E., Feílix, C., Ferreira, E. (2010). Evaluation of hollocelulase production by plant-degrading fungi grown on agro-industrial residues. Biodegradation 21, 815-824.         [ Links ]

Gordillo, F., Caputo, V., Peirano, A., Chavez, R., van Beeumen, J., Vandenberghe I., Claeyssens, M., Bull, P., Ravanal, M., Eyzaguirre, J. (2006). Penicillum purpurogenum produces a family 1 acetyl xylan esterase containing a carbohydrate binding domain: characterization of the protein and its gene. Micologycal Research 110, 1129-1139.         [ Links ]

Graham, H., Hesselman, K., Aman, A. (1986). The influence of wheat bran and sugar-beet pulp on the digestibility of dietary components in a cereal-based pig diet. The Journal of Nutrition 116, 242-251.         [ Links ]

Kabel, M., Borne, H., Vincken, J.P., Voragen, A.G.J., Schols, H. (2007). Structural differences of xylans affect their interaction with cellulose. Carbohydrate Polymers 69, 94-105.         [ Links ]

Kadam, K., McMillan, J. (2003). Availability of corn stover as a sustainable feedstock for bioethanol production. Bioresource Technology 88, 17-25.         [ Links ]

Kiyohara, M., Sakaguchi, K., Yamaguchi, K., Araki, T., Ito, M. (2009). Characterization and application of carbohydrate-binding modules of b-1,3 xylanase XYL 4. The Journal of Biochemistry146, 633-641.         [ Links ]

Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685.         [ Links ]

Luis, A., Venditto, I., Temple, M., Rogowsky A., Basleí, A., Xu, J., Know, P., Prates, J., Ferreira, L., Fontes, C., Najmudin, S., Gilbert, H. (2013). Understanding how non catalytic carbohydrate binding modules can display specificity for xyloglucan. The Journal of Biological Chemistry 288,4799- 4809.         [ Links ]

Miller, G.L. (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry 31, 426-428.         [ Links ]

Mogollón, G., García, J.A., León, W. (2008). Materias primas fibrosas. En: Panorama de la industria de celulosa y papel en Iberoameírica (Ma.C. Areaedit), Pp 15-60. Red Iberoamericana de docencia e investigacioín en celulosa y papel, Argentina.         [ Links ]

Ng, T.B. (2004). Peptides and Proteins from Fungi. Peptides 25, 1055-1073.         [ Links ]

Nigam, P., Pandey, A. (2009). Solid state fermentation technology for bioconversion of biomass of agricultural residues. En: Biotechnology for Agro industrial Residues Utilization (P.S. Nigam, A. Pandey eds.), Pp 197-221. Springer Science + Bussines Media.         [ Links ]

Okeke, B.C., Obi, S.K. (1994). Lignocellulose and sugar composition of some agro-waste materials. Bioresource Technology 47, 283-284.         [ Links ]

Paldi, T., Levy, I., Shosheyov, O. (2003). Glucoamylase starch binding-domain in Aspergillus niger B1: molecular cloning and functional characterization. Biochemistry Journal 372, 905-910.         [ Links ]

Quentin, M., Ebbelaar, M., Derksen, J., Mariani, C., van der Valk, H. (2002). Description of a cellulose-binding domain and a linker sequence from Aspergillus fungi. Applied Microbial Biotechnology 58, 658-662.         [ Links ]

Quiquampoix, H. (2002). Enzyme adsorption on soil mineral surfaces and consequences for the catalytic activity. En: Enzymes in the Environment. Activity, Ecology and Applications (R. Burns and R. Dick eds.) Pp 285-306. Marcel Dekker Inc, New York.         [ Links ]

Van Tassel, P., Viot, P., Tarjus, G. (1997). A kinetic model of partially reversible protein adsorption. Journal of Chemical Physical 106, 961-770.         [ Links ]