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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This research was supported by the Spanish Ministry of Education and Science (RYC-2007-01667, PPQ2006-08230, CONSOLIDER-C BIO2006 05668, and CONSOLIDER-CSD 2007-00055). Araceli Crespo and Isabel Jimenez are also gratefully acknowledged for their practical assistance.
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Muñoz, R., Hernández, M., Segura, A. et al. Continuous cultures of Pseudomonas putida mt-2 overcome catabolic function loss under real case operating conditions. Appl Microbiol Biotechnol 83, 189–198 (2009). https://doi.org/10.1007/s00253-009-1928-5
- Microbial damage
- Plasmid pWW0
- Process stability
- Pseudomonas putida mt-2
- Solvent tolerance
- Toluene biodegradation