Correo Electrónico
DNINFOA - SIA
Bibliotecas
Convocatorias
Identidad U.N.
Published
Evaluating the effect of the number of generations in IBUN 91.2.98 Leuconostoc mesenteroides cultures on enzyme extract production
Evaluación del efecto del número de generaciones en un cultivo de Leuconostoc Mesenteroides IBUN 91.2.98 sobre la producción de un extracto enzimático
Keywords:
Leuconostoc mesenteroides, generation number, scale-up, glucosyltransferase activity. (en)Leuconostoc mesenteroides, número de generaciones, escalado, actividad glucosiltransferasa. (es)
This work studied the effect of the number of generations of the IBUN 91.2.98 Leuconostoc mesenteroides strain on enzyme complex production. The subculturing technique was used on a medium which had been designed specifically for this organism for producing an enzyme complex. The effect was indirectly determined by monitoring microorganism growth and measuring the glucosyltransferase and hydrolytic activity of an enzyme extract obtained from such culture. There were 40 subcultures, representing 196 generations of IBUN 91.2.98 Leuconostoc mesenteroides. The results led to establishing that the extract's higher enzymatic activity (from 4 to 6 U/mL) was reached at the end of the culture's exponential phase and that this activity was stable during subculturing, confirming that there was no variation in strain regarding enzyme extract production until such number of generations had occurred, thereby not being limited to scaling-up to 8,000 litres.
Por medio de este trabajo de investigación se estudió el efecto del número de generaciones de la cepa IBUN 91.2.98 de Leuconostoc mesenteroides sobre la producción de un complejo enzimático. Para ello se empleó la técnica de subcultivos en un medio diseñado específicamente para este microorganismo, con fines de producción de un complejo enzimático. La estabilidad se determinó, de manera indirecta, por medio del seguimiento del crecimiento del microorganismo y la medición de las actividades glucosiltransferasa e hidrolítica de un extracto enzimático obtenido del cultivo. En total se realizaron 40 subcultivos, correspondientes a 196 generaciones de Leuconostoc mesenteroides IBUN 91.2.98. Los resultados obtenidos permiten establecer que la mayor actividad del extracto enzimático (entre 4 y 6 U/mL) se alcanza al final de la fase exponencial del cultivo y que dicha actividad se mantiene estable durante los subcultivos, confirmando que hasta ese número de generaciones no se presenta una variación de la cepa en la producción del extracto enzimático, por lo que no constituye límite para escalar hasta 8.000 litros.
Downloads
References
Breton, C., Snajdrová, L.Š., Jeanneau, C., Koca, J., Imberty, A. Structures and mechanisms of glycosyltransferases. Glycobiology, Vol. 16, No.2, 2006, pp. 29 - 37.
Cardozo, A., Gómez, M., Aislamiento e identificación de cepas nativas con actividad levansacarasa, BSc Bacteriology thesis, Universidad Colegio Mayor de Cundinamarca, 1998.
Cuesta, B. Aseguramiento de Calidad. Cultivos de referencia. Norma ISO 17025: 1-44.
Dols, S., Willemot, R. M., Vignon, M., Monsan, P., Characterization of the different dextransucrase activities excreted in glucose, fructose, or sucrose medium by Leuconostoc mesenteroides NRRL B-1299., Applied and Environmental Microbiology, Vol.64, No.4, Abril, 1998, pp.1298-302.
Dols, M., Remaud-Simeon, M., Monsan, P. F., Dextransucrase production by Leuconostoc mesenteroides NRRL B-1299, comparison with L. mesenteroides NRRL B-512F., Enzyme Microbiology Technology. Vol 20,1997, pp.523-530.
García, M., Villamizar, L., Torres L., Cotes A., Efecto de subcultivos sucesivos de Beauveria bassiana sobre sus características y actividad contra Premnotrypes voraz. Manejo Integrado de Plagas y Agroecología. Vol. 77, 2006, pp. 50-56.
Giraldo, S. E., Mora, C., Ospina, S. Caracterización de las propiedades de hidrofilidad de levana de origen biotecnológico. Primer Congreso Colombiano de Biotecnología,. Bogotá,. Universidad Nacional de Colombia, 2002, pp.160-160 v.
Kaneko, Y., Sato, R., Aoyagi, H., Evaluation of Chinese hamster ovary cell stability during repeated culture for large scale antibody production. Journal of Bioscience and Bioengineering. Vol. 109, No. 3, 2010, pp. 274-280.
Kobayashi, M., K. Matsuda. Purification and characterization of two activities of the intracellular dextransucrase from Leuconostoc mesenteroides NRRL B-1299. Biochim Biophys Acta. Vol. 397, 1975, pp.69-79.
Miller, G. L. Use of Dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry. Vol 31,1959, pp. 426-428.
Moulis, C., Joucla, G., Harrison, D., Fabre, E., Monsan, P., Understanding the polymerization mechanism of glycoside-hydrolase family 70 glucansucrases. Journal of Biological Chemistry, Vol. 281 No. 42, Oct.,2006, pp. 31254-67.
Purama, R. K., Goyal, A. Identification, effective purification and functional characterization of dextransucrase from Leuconostoc mesenteroides NRRL B-640. Bioresource Technology., 2008 pp. 3635-3642.
Saenz, A. C., Prado, M., Aislamiento de cepas nativas con actividad levansacarasa, BSc Bacteriology thesis, Universidad Colegio Mayor de Cundinamarca, 1998.
Soler, A., Buitrago, H. Evaluación de la transferencia de oxígeno en cultivos con Lactococcus lactis empleando un sistema de fermentación con aireación externa. Revista Colombiana de Biotecnología. Vol 12, No.2, 2010, pp.124-138.
Thiry, M., Cingolani, D., Optimizing scale-up fermentation process. Trends in Biotechnology. Vol. 3, 2002, pp.103-105.
License
Copyright (c) 2013 Gustavo Buitrago Hurtado, Withney Andrea Villamil Porras, Daissy Jasbeydi Vargas Sepúlveda, Ángela Otálvaro Alvarez, Glaether Yohn Flórez
This work is licensed under a Creative Commons Attribution 4.0 International License.
The authors or holders of the copyright for each article hereby confer exclusive, limited and free authorization on the Universidad Nacional de Colombia's journal Ingeniería e Investigación concerning the aforementioned article which, once it has been evaluated and approved, will be submitted for publication, in line with the following items:
1. The version which has been corrected according to the evaluators' suggestions will be remitted and it will be made clear whether the aforementioned article is an unedited document regarding which the rights to be authorized are held and total responsibility will be assumed by the authors for the content of the work being submitted to Ingeniería e Investigación, the Universidad Nacional de Colombia and third-parties;
2. The authorization conferred on the journal will come into force from the date on which it is included in the respective volume and issue of Ingeniería e Investigación in the Open Journal Systems and on the journal's main page (https://revistas.unal.edu.co/index.php/ingeinv), as well as in different databases and indices in which the publication is indexed;
3. The authors authorize the Universidad Nacional de Colombia's journal Ingeniería e Investigación to publish the document in whatever required format (printed, digital, electronic or whatsoever known or yet to be discovered form) and authorize Ingeniería e Investigación to include the work in any indices and/or search engines deemed necessary for promoting its diffusion;
4. The authors accept that such authorization is given free of charge and they, therefore, waive any right to receive remuneration from the publication, distribution, public communication and any use whatsoever referred to in the terms of this authorization.
