Producción de proteína y biomasa probiótica de Lactobacillus casei liofilizadas a partir de suero de leche de cabra

Production of freeze dried protein and Lactobacillus casei probiotic biomass from goat milk whey

E. J. Aguirre–Ezkauriatza, A. Ramirez–Medrano, J. M. Aguilar–Yáñez y M. M. Alvarez*

Centro de Biotecnología–FEMSA del Tecnológico de Monterrey Av. Eugenio Garza Sada 2501 sur. Col. Tecnológico. Edificio CB, 5to piso. Monterrey, N.L. 64849, México. * Autor para la correspondencia. E–mail: mario.alvarez@itesm.mx Tel: (81) 83284131, Fax: 83284136

Resumen

En este trabajo se documenta la factibilidad técnica de obtención de dos productos de valor agregado a partir de suero de leche de cabra: (a) proteína liofilizada y (b) biomasa probiótica liofilizada de Lactobacillus casei. Se presentan resultados de experimentos de ultrafiltración de suero de leche de cabra (SLC), del cual se retiene proteína con una alta relación proteína/lactosa. En su forma liofilizada, esta proteína presentó características de solubilidad superiores a productos comerciales similares. El permeado resultante del ultrafiltrado de SLC fue utilizado como medio de cultivo para el crecimiento del microorganismo probiótico L. casei. Se reportan resultados de experimentos de fermentación de suero de leche por lote y lote alimentado para la producción de biomasa. La cinética de crecimiento de L. casei en SLC desproteinizado no suplementado es analizada a partir de datos de experimentos por lotes. Se observa un marcado efecto inhibitorio del crecimiento bacteriano asociado a la producción de ácido láctico. Las estrategias por lote alimentado aquí reportadas resultan en productividades superiores, concentraciones de sustrato residual menores y viabilidades más altas del producto respecto a procesos por lote.

Palabras clave: Lactobacillus casei, por lote alimentado, biorreactores, suero de leche, cabra, probiótico.

Abstract

In this communication, we document the technical feasibility of elaboration of two added value products from goat whey: (a) freeze–dried protein and (b) freeze–dried probiotic biomass of Lactobacillus casei. Results from goat milk whey ultrafiltration experiments are presented, from which high protein/lactose ratio protein is recovered. In its lyophilized form, this protein presents superior solubility characteristics with respect to analogous commercial products. The ultrafiltration permeate was utilized as growth culture medium for the probiotic microorganism L. casei. Results from batch and fed–batch fermentation experiments are reported. The growth kinetics of L. casei in de–proteinized goat milk whey is analyzed. From batch experiments, a kinetic characterization is conducted. A strong inhibition of biomass growth by lactic acid production is observed. Fed–batch strategies resulted in higher average productivities, lower residual substrate concentrations, and higher product viable counts than batch strategies.

Keywords: Lactobacillus casei, fed–batch, bioreactors, goat milk, whey, probiotic.

DESCARGAR ARTÍCULO EN FORMATO PDF

Agradecimientos

Agradecemos a la empresa Caprico® por suministrar el suero de leche utilizado en los experimentos aquí reportados. Agradecemos el financiamiento provisto por el Tecnológico de Monterrey, a través del fondo de Investigación CAT122.

Referencias

Alvarez, M.M. y Aguirre–Ezkauriatza, J.E. (2008). Proceso de alto rendimiento para la producción de biomasa probiótica de bacterias lácticas a partir de suero de leche no suplementado. Patente mexicana en trámite. MX/a/2008/016564.         [ Links ]

Aguirre–Eskauriatza, J.E. (2008). Producción de proteína liofilizada y biomasa de Lactobacillus casei BPG4 liofilizada, a partir de suero de leche de cabra. Tesis de Maestría. Tecnológico de Monterrey, México.         [ Links ]

Altiok, D., Tokatli, F. y Harsa, S. (2006). Kinetic modeling of lactic acid production from whey by Lactobacillus casei (NRRL B–441). Journal of Chemical Technoogy and Biotechnology 81,1190–1197.         [ Links ]

Brink, M. y Todorov, S.D. (2006). The effect of probiotics on production of antimicrobial compounds, resistance to growth at low pH and in the presence of bile, and adhesion of probiotic cells to intestinal mucus. Journal of Applied Microbiology 100(4), 813–820.         [ Links ]

De Man, J.C., Rogosa, M. y Sharpe, M.E. (1960). A medium for the cultivation of Lactobacilli. Journal of Applied Microbiology 23(1),130– 135.         [ Links ]

Fairbrother, P., George, W.O. y Williams, J.M. (1991). Whey fermentation: on–line analysis of lactose and lactic acid by FTIR spectroscopy. Applied Microbiology & Biotechnology 35,301–305.         [ Links ]

Gaon, D. y Doweck, Y. (1995). Lactose digestion by milk fermented with Lactobacillus acidophilus and Lactobacillus casei of human origin. Medicina (B Aires) 55(3),237–42.         [ Links ]

Ha, M.Y. y Kim, S.W. (2003). Kinetics analysis of growth and lactic acid production in pH–controlled batch cultures of Lactobacillus casei KH–1 using yeast extract/corn steep liquor/glucose medium. Journal of Bioscience and Bioengineering 96(2),134–140.         [ Links ]

Haarman, M. y Knol, J.(2006). Quantitative real–time PCR analysis of fecal Lactobacillus species in infants receiving a prebiotic infant formula. Applied Environmental Microbiology 72(4), 2359–2365.         [ Links ]

Hayakawa, K., Harada, K., Takeuchi, S., Shibata, S. y Miyagi, A. (2000). "Method for controlling culture of lactic bacteria." US Patent 6054262. Issued April 25, 2000.         [ Links ]

Ingrassia, I. y Leplingard, A. (2005). Lactobacillus casei DN–114 001 inhibits the ability of adherent–invasive Escherichia coli isolated from Crohn's disease patients to adhere to and to invade intestinal epithelial cells. Applied Environmental Microbiology 71(6), 2880–2887.         [ Links ]

Isolauri, E. (1991). A human Lactobacillus strain (Lactobacillus casei sp. strain GG) promotes recovery from acute diarrhea in children. Pediatrics 88(1), 90–97.         [ Links ]

Isolauri, E. y Kaila, M. (1993). Diet during rotavirus enteritis affects jejunal permeability to macromolecules in suckling rats. Pediatric Research 33(6), 548–553.         [ Links ]

Liong, M.T. y Shah, N.P. (2006). Effects of a Lactobacillus casei synbiotic on serum lipoprotein, intestinal microflora, and organic acids in rats. Journal of Dairy Science 89(5), 1390–1399.         [ Links ]

Mondragón–Parada, M.E. y Nájera–Martínez, M. (2006). Lactic acid bacteria production from whey. Applied Biochemistry and Biotechnology 134(3), 223–32.         [ Links ]

Nemeth, E. y Fajdiga, S. (2006). Inhibition of Salmonella–induced IL–8 synthesis and expression of Hsp70 in enterocyte–like Caco–2 cells after exposure to non–starter lactobacilli. International Journal of Food Microbiology 112(3), 266–274.         [ Links ]

Nerlich, B. y Koteyko, N. (2008). Balancing food risks and food benefits: the coverage of probiotics in the UK national press. Sociological Research Online 13(3).         [ Links ]

Ortiz–Andrellucchi, A. y Sánchez–Villegas, A. (2008). Immunomodulatory effects of the intake of fermented milk with Lactobacillus casei DN114001 in lactating mothers and their children. British Journal of Nutrition 1,1–12.         [ Links ]

Pham, M., Lemberg, D.A. y Day, A.S. (2008). Probiotics: sorting the evidence from the myths. Medical Journal of Australia 188(5), 304–308.         [ Links ]

Parassol, N. y Freitas, M. (2005). Lactobacillus casei DN–114 001 inhibits the increase in paracellular permeability of enteropathogenic Escherichia coli infected T84 cells. Research in Microbiology 156(2), 256–62.         [ Links ]

Park, Y.H. y Kim, J.G. (2008). Effects of Lactobacillus acidophilus 43121 and a mixture of Lactobacillus casei and Bifidobacterium longum on the serum cholesterol level and fecal sterol excretion in hypercholesterolemia–induced pigs. Bioscience, Biotechnology and Biochemistry 72(2), 595–600.         [ Links ]

Soto–Cruz, O. y Páez–Lerma, J. (2005). Balances en procesos de fermentación: análisis de consistencia y de flujos metabólicos (Fermentation process balances: consistency and metabolic flux analysis). Revista Mexicana de Ingeniería Química 4, 59–74.         [ Links ]

Srinivasan, R. y Meyer, R. (2006). Clinical safety of Lactobacillus casei strain shirota as a probiotic in critically ill children. Journal of Pediatric Gastroenterology and Nutrition 42(2), 171–173.         [ Links ]

Rosmini, M.R., Sequeira, G. J., Guerrero–Legarreta, I., Martí, L. E., Dalla–Santina, R., Frizzo, L. y Bonazza J.C. (2004). Producción de probióticos para animales de abasto: importancia del uso de la microbiota intestinal indígena (Probiotic production for meat animals: importance of using indigenous intestinal microbiota). Revista mexicana de ingenieria quimica 3, 181–191.         [ Links ]

Youssef, C.B. y Goma, G. (2005). Kinetic modeling of Lactobacillus casei ssp. rhamnosus growth and lactic acid production in batch cultures under various medium conditions. Biotechnology Letters 27(22), 1785–1789.         [ Links ]

Yuki, N., Watanabe, K., Mike, A., Tagami, Y., Tanaka, R., Ohwaki, M. y Morotomi, M. (1999). Survival of a probiotic, Lactobaillus casei strain Shirota in the gastrointestinal tract: selective isolation from feces and identification using monoclonal antibodies. International Journal of Food Microbiology 48(1), 51–57.         [ Links ]