Continuous cultures of Pseudomonas putida mt-2 overcome catabolic function loss under real case operating conditions |
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Authors: | Raúl Muñoz María Hernández Ana Segura Joao Gouveia Antonia Rojas Juan Luis Ramos Santiago Villaverde |
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Affiliation: | (1) Department of Chemical Engineering and Environmental Technology, Valladolid University, Paseo del Prado de la Magdalena, s/n, Valladolid, Spain;(2) Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Profesor Albareda, 1, Granada, 18008, Spain;(3) Calantia Biotech, CEEI Valencia/Parque Tecnológico, Avd. Benjamin Franklin 12, 46980 Paterna, Valencia, Spain |
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Abstract: | The long-term performance and stability of Pseudomonas putida mt-2 cultures, a toluene-sensitive strain harboring the genes responsible for toluene biodegradation in the archetypal plasmid pWW0, was investigated in a chemostat bioreactor functioning under real case operating conditions. The process was operated at a dilution rate of 0.1 h−1 under toluene loading rates of 259 ± 23 and 801 ± 78 g m−3 h−1 (inlet toluene concentrations of 3.5 and 10.9 g m−3, respectively). Despite the deleterious effects of toluene and its degradation intermediates, the phenotype of this sensitive P. putida culture rapidly recovered from a 95% Tol− population at day 4 to approx. 100% Tol+ cells from day 13 onward, sustaining elimination capacities of 232 ± 10 g m−3 h−1 at 3.5 g Tol m−3 and 377 ± 13 g m−3 h−1 at 10.9 g Tol m−3, which were comparable to those achieved by highly tolerant strains such as P. putida DOT T1E and P. putida F1 under identical experimental conditions. Only one type of Tol− variant, harboring a TOL-like plasmid with a 38.5 kb deletion (containing the upper and meta operons for toluene biodegradation), was identified. |
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Keywords: | Microbial damage Plasmid pWW0 Process stability Pseudomonas putida mt-2 Solvent tolerance Toluene biodegradation |
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