Adjustment and Scale-Up Strategy of Pilot Liquid Fermentation Process of Azotobacter sp.
The genus Azotobacter has been widely used as bio-fertilizer due to its significant effects on the stimulation and promotion of plant growth in various agricultural species of commercial interest. In order to obtain significantly viable cellular concentration, a scale-up strategy for a liquid fermentation process (SmF) with two strains of A. chroococcum (named Ac1 and Ac10) was validated and adjusted at laboratory and pilot scale. A batch fermentation process under previously defined conditions was carried out on a biorreactor Infors®, model Minifors of 3.5 L, which served as a baseline for this research. For the purpose of increasing process efficiency, the effect of the reduction of stirring speed was evaluated in combination with a fed-batch-type fermentation laboratory scale. To reproduce the efficiency parameters obtained, a scale-up strategy with geometric and fluid dynamic behavior similarities was evaluated. According to the analysis of variance, this scale-up strategy did not have significant effect on cellular concentration and in laboratory and pilot fermentations (Tukey, p > 0.05). Regarding air consumption, fermentation process at pilot scale showed a reduction of 23% versus the baseline. The percentage of reduction related to energy consumption reduction under laboratory and pilot scale conditions was 96.9% compared with baseline.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1130353Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 756
 C. Ervin, D. Ervin, “Factors Affecting the use of soil conservation practices: Hypotheses, Evidence, and Policy Implications,” Land Economics, vol. 58, no. 3, pp. 272-292, Aug. 1982.
 E. Lutz, S. Pagiola, C. Reiche, “The costs and benefits of soil conservation: The farmer´s Viewpoint,” The Word Bank Research Observer, vol. 9, no. 2, pp. 273-295, July, 1994.
 M. Yussefi, H. Willer, “Organic Farming Worldwide 2007: Overview and main statistics,” in The world of organic agriculture, Bonn: IFOAM & FiBL, 2007, pp. 9-22.
 C. Cruz, L. Barrero, F. Rodríguez, “Analysis of bioprospecting processes in Colombia,” in Bioprospecting for the development of the agricultural sector of Colombia, Mosquera, 2012, pp. 21-30.
 M. Camelo, “Technological development of a biofertilizer based on the diazotrophic bacterium Azotobacter chroococcum,” Master’s Thesis, Military University Nueva Granada, Bogotá, 2010, pp. 8-13.
 N. Mrkovacki, V. Millic, “Use of Azotobacter chrocooccum as potentially useful in agricultural applications,” Annals of Microbiol., vol. 51, pp. 145-158, 2001
 M. Brown, “Role of Azotobacter paspaliin Association with Paspalum notatum,” J. Applied Bacteriol., vol. 40, no. 3, pp. 341-348, June 1976
 A. Moreno-Galván, D. Rojas, R. Bonilla, “Sequential statistical design application in identification of Azotobacter chroococcum AC1 nutritional sources,” J. CORPOICA - Agricultural Science and Technology, vol. 11, no. 2, pp. 151-158, Nov. 2011
 A. Lara, L. Palomares, O. Ramírez, “Bioreactor scale up,” in Encyclopedia of cell Technol., 2000, pp. 1-22
 J. Martinko, M. T. Madigan, K. S. Bender, D. H. Buckley, D. A. Stahl, T. Brock, Brock: biology of microorganisms, Madrid: Pearson, 12th edition, 2009.
 J. Chimero, “Estudio sobre la depuración de los lixiviados de RSU con cenizas volátiles zeolitizadas,” Master’s Thesis, 2006.
 G. L. Turner, A. H. Gibson, “Measurement of nitrogen fixation by indirect means,” in Methods for Evaluating Biological Nitrogen Fixation, Wiley, Ed., Chichester, 1980. pp. 111–139.
 A. Díaz-Barrera, A. Aguirre, J. Berrios, F. Acevedo, “Continuous cultures for alginate production by Azotobacter vinelandii growing at different oxygen uptaken rates,” Process Biochemistry, vol. 46, no. 9, pp. 1879-1883, June 2011.
 C. Then, Z. Othman, W. Mustapha, M. Sarmidi, R. Aziz, H. El Enshasy, “Production of alginate by Azotobacter vinelandii in semi-industrial scale using batch and fed-batch cultivations systems,” J. of Adv. Scien. Research, vol. 3, no. 4, pp. 45-50, Jan. 2012.
 M. Doyle, L. R. Beuchat, T. J. Montville, Microbiología de los alimentos. Fundamentos y fronteras, 2ª ed., España: Acribia Ed., 2001, pp. 773-785.
 R. M. Maier, I. L. Pepper, C. P. Gerba, “Bacterial Growth,” in Environmental Microbiology, 3rd ed., San Diego, CA: Academic Press Inc, 2009, pp. 40.
 R. Perry, Handbook of Chemical Engineering, 4th ed., McGraw-Hill Ed., 2001.
 P. Harriott, “Nonideal Flow” in Chemical Reactor Design. 1st ed. New York: CRC Press Ed., 2003, pp. 231-262.
 A. Anaya, H. Pedroza, “Scale-up, the art of chemical engineering: Pilot plants the passage between the egg and the hen,” Technology, Science, Education, vol. 23, no. 1, pp. 31-39, 2008.
 C. Hewitt, A. Nienow, “The scaleup of microbial in batch and fedbatch fermentation processes,” Advances in Applied Microbiology, vol. 62, pp. 105-135, 2007.
 F. Schmidt, “Optimization and scale up of industrial fermentation processes,” Appl. Microbiol Biotechnol, vol. 68, no. 4, pp. 425-435, Oct. 2005.
 F. Clementi, “Alginate production by Azotobacter vinelandii”, Crit. Reviews in Biotechnol., vol. 17, no. 4, pp. 327-361, 1997.
 M. Mejía, D. Segura, G. Espín, E. Galindo and C. Peña, “Two-stage fermentation process for alginate production by Azotobacter vinelandii mutant altered in poly-β-hydroxybutyrate (PHB) synthesis”, J. of Applied Microbiol, vol. 108, pp. 55-61, April 2009.
 O. Damir, M. Pavlecic, B. Santek, S. Novak, “Cultivation of the bacterium Azotobacter chroococcum for preparation of biofertilizers,” African Journal of Biotechnology, vol. 10, no. 16, pp. 3104-3111, April 2011.
 C. Pozo, M. Martínez-Toledo, B. Rodelas, J. González-López, “Effects of culture conditions on the production of polyhydroxyalkanoates by Azotobacter chroococcum H23 in media containing a high concentration of alpechin as primary carbon source,” J. of Biotechol., vol. 97, no. 2, pp. 125-131, 2002.
 C. Reyes, C. Peña, E. Galindo, “Reproducing shake flask performance in stirred fermentors: production of alginates by Azotobacter vinelandii,” J. of Biotechol., vol. 105, no. 1-2, pp. 189-198, Nov. 2003.
 C. Peña, M. Millán, E. Galindo, “Production of alginate by Azotobacter vinelandii in a stirred fermentor simulating the evolution of power input observed in shake flasks,” Process Biochemistry, vol. 43, pp. 775-778, July 2008.
 B. Juárez, J. Martínez-Toledo, J. González-López, “Growth of Azotobacter chroococcum in chemically defined media containing p-hydroxybenzoic acid and protocatechuic acid,” Chemosphere, vol. 59, no. 3, pp. 136-1365, Dec. 2001.
 R. Quintero, Biochemical Engineering. Theory and applications, Mexico: Ediciones Alhambra Mexicana, 1981.
 L. Maranga, A. Cunha, J. Clemente, P. Cruz, M. Carrondo, “Scale‐up of virus‐like particles production: effects of sparging, agitation and bioreactor scale on cell growth, infection kinetics and productivity,” J Biotechnol, vol. 107, no. 1 pp. 55‐64, Jan. 2004.