Abstract
Azospirillum brasilense has industrial significance as a growth promoter in plants of commercial interest. However, there is no report in the literature disclosing a liquid product produced in pilot-scale bioreactors and is able to be stored at room temperature for more than 2 years. The aim of this work was to scale up a process from a shake flask to a 10-L lab-scale and 1,000-L pilot-scale bioreactor for the production of plant growth-promoting bacterium A. brasilense for a liquid inoculant formulation. Furthermore, this work aimed to determine the shelf life of the liquid formulation stored at room temperature and to increase maize crops yield in greenhouses. Under a constant oxygen mass transfer coefficient (K L a), a fermentation process was successfully scaled up from shake flasks to 10- and 1,000-L bioreactors. A concentration ranging from 3.5 to 7.5 × 108 CFU/mL was obtained in shake flasks and bioreactors, and after 2 years stored at room temperature, the liquid formulation showed one order of magnitude decrease. Applications of the cultured bacteria in maize yields resulted in increases of up to 95 % in corncobs and 70 % in aboveground biomass.
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Acknowledgments
This work was partially financed by the Consejo Nacional de Ciencia y Tecnología (CONACyT grants 178528, CONACYT-INNOVAPYME 181895), and the Programa de Apoyo a Proyectos de Investigación e Innovación Tecnológica, Universidad Nacional Autónoma de México (PAPIIT-UNAM IN-210013 and IN-209113). Authors thanks Ce Akatl Mikiztli Arias-Poblano, Food Chem.; Monserrat García-García, Biol.; Maria Fernanda Nava-Ocampo, Chem. Eng.; Angeles Cancino-Rodezno, Ph.D.; David Fernando Zuluaga-Rave, M.Sc.; Andrea Bedoya-Lopez, M.Sc.; Andrea Castellanos-Mendoza, Pharm. Biol. Chem.; Pedro Navarro-Garcia, Pharm. Biol. Chem.; Luís Gutiérrez, Food Chem.; Blanca Garnica-Garcia, Food Chem.; Liliana Guadalupe Vigueras, Pharm. Biol. Chem.; and Ingue Karen Carrasco-Espinosa, Biol. (Bioprocess Unit staff, Instituto de Investigaciones Biomédicas, UNAM), for their assistance with the 1,000-L cultures. LDMP thanks the scholarship from Colciencias, Colombia. RAGS thanks the scholarship from CONACYT, México. The authors thank Marcel Morales, Ph.D. (CEO of Biofábrica Siglo XXI S.A. de C.V.) and Agustin de Leonardo, Eng. (process manager of Biofábrica Siglo XXI S.A. de C.V.). Technical assistance on bioreactor controllability by Dusstthon Llorente, Eng. is also appreciated. We thank Dr. Martha G. López-Guerrero for her critical review of the manuscript. We also thank Ana Carmen Delgado for reviewing the English version of the manuscript.
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The authors declare that they have no conflict of interest.
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This study is dedicated to the memory of Professor Jesus Caballero-Mellado, Ph.D. (1953–2010) of the Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, the pioneer of Azospirillum research in Mexico.
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Trujillo-Roldán, M.A., Valdez-Cruz, N.A., Gonzalez-Monterrubio, C.F. et al. Scale-up from shake flasks to pilot-scale production of the plant growth-promoting bacterium Azospirillum brasilense for preparing a liquid inoculant formulation. Appl Microbiol Biotechnol 97, 9665–9674 (2013). https://doi.org/10.1007/s00253-013-5199-9
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DOI: https://doi.org/10.1007/s00253-013-5199-9