, Volume 19, Issue 6, pp 897–908 | Cite as

Response of Pseudomonas putida F1 cultures to fluctuating toluene loads and operational failures in suspended growth bioreactors

  • Raúl Muñoz
  • Luís Felipe Díaz
  • Sergio Bordel
  • Santiago Villaverde
Original Paper


The response of Pseudomonas putida F1 to process fluctuations and operational failures during toluene biodegradation was evaluated in a chemostat suspended growth bioreactor. The ability of P. putida F1 to rapidly increase its specific toluene degradation capacity resulted in no significant variation in process removal efficiency when toluene load was increased from 188 to 341 g m−3 h−1. Likewise, bacterial activity rapidly reached steady state performance (in less than 1.5 h after the restoration of steady state operational conditions) following an 8 h process shutdown, or after episodes of toluene or mineral nutrients deprivation. Process performance was however highly sensitive to pH, as pH levels below 4.5 dramatically inhibited bacterial activity, decreasing severely process robustness and inducing a cycle of periodic process collapses and recoveries. This pH mediated deterioration of bacterial activity was confirmed by further respirometric tests, which revealed a 50–60% reduction in the O2 consumption rate during the degradation of both toluene and 3-methyl catechol when pH decreased from 5.05 to 4.55. Finally, process robustness was quantified according to methods previously described in literature.


Gas treatment Pseudomonas putida F1 Robustness Suspended growth bioreactors VOC biodegradation 



This research was supported by the Spanish Ministry of Education and Science (PPQ2006-08230 and RYC-2007-01667).


  1. Baquerizo G, Maestre JP, Gamisans X, Gabriel D, Lafuente J (2007) Study of NH3 removal by Gas-Phase Biofiltration: effects of shock loads and watering rate on biofilter performance. In: Proceedings of the II international congress on biotechniques for air pollution control, A Coruña, Spain, 3–5 October 2007Google Scholar
  2. Barona A, Elias A, Arias R, Cano R, González R (2004) Biofilter response to gradual and sudden variations in operating conditions. Biochem Eng J 22:25–31CrossRefGoogle Scholar
  3. Bordel S, Muñoz R, Díaz LF, Villaverde S (2007) New insights on toluene biodegradation by Pseudomonas putida F1: influence of pollutant concentration and excreted metabolites. Appl Microbiol Biotechnol 74:857–866PubMedCrossRefGoogle Scholar
  4. Boudreau NG, Daugulis AJ (2006) Transient performance of two-phase partitioning bioreactors treating a toluene contaminated gas stream. Biotechnol Bioeng 94:448–457PubMedCrossRefGoogle Scholar
  5. Diaz LF, Muñoz R, Bordel S, Villaverde S (2008) Toluene biodegradation by Pseudomonas putida F1: targeting culture stability in long term operation. Biodegradation 19(2):197–202PubMedCrossRefGoogle Scholar
  6. Elena SF, Lenski RE (2003) Microbial genetics: evolution experiments with microorganisms: the dynamics and genetic bases of adaptation. Nat Rev Genet 4:457–469PubMedCrossRefGoogle Scholar
  7. Halecky M, Paca J, Gerrard AM, Soccol CR (2007) Styrene degradation in perlite biofilter: the overall performance characteristics and dynamic response. In: Proceedings of the II international congress on biotechniques for air pollution control, A Coruña, Spain, 3–5 October 2007Google Scholar
  8. Jenkins RO, Heald SC (1996) Stability of toluene oxidation by Pseudomonas putida under nutrient deprivation. Appl Microbiol Biotechnol 46:388–392CrossRefGoogle Scholar
  9. Jung IG, Park OH (2005) Enhancement of cometabolic biodegradation of trichloroethylene (TCE) gas in biofiltration. J Biosci Bioeng 100:657–661PubMedCrossRefGoogle Scholar
  10. Kennes C, Thalasso F (1998) Waste gas biotreatment technology. J Chem Technol Biotechnol 72:303–319CrossRefGoogle Scholar
  11. Kraakman NJR (2003) Robustness of a full-scale biological system treating industrial CS2 emissions. Environ Prog 22:79–85CrossRefGoogle Scholar
  12. Kraakman NJR (2005) Biotrickling and bioscrubbers applications to control odor and air pollutants: developments, implementation issues and case studies. In: Shareefdeen Z, Singh A (eds) Biotechnology for odour and air pollution control. Springer-Verlag, HeidelbergGoogle Scholar
  13. Maestre JP, Gamisans X, Gabriel D, Lafuente J (2007) Fungal biofilters for toluene biofiltration: evaluation of the performance with tour parking materials under different operating conditions. Chemosphere 67:684–692PubMedCrossRefGoogle Scholar
  14. Metris A, Gerrard AM, Cumming RH, Weigner P, Paca J (2001) Modelling shock loadings and starvation in the biofiltration of toluene and xylene. J Chem Technol Biotechnol 76:565–572CrossRefGoogle Scholar
  15. Moe WM, Qi B (2004) Performance of a fungal biofilter treating gas-phase solvent mixtures during intermittent loading. Water Res 38:2259–2268CrossRefGoogle Scholar
  16. Morales M, Velázquez E, Jan J, Revah S, González U, Razo-Flores E (2004) Methyl tert-butyl ether biodegradation by microbial consortia obtained from soil samples of gasoline-polluted sites in Mexico. Biotechnol Lett 26:269–275PubMedCrossRefGoogle Scholar
  17. Nielsen DR, Sak KN, Mclellan PJ, Daugulis AJ (2006) Benzene vapor treatment using a two-phase partitioning bioscrubbers: an improved steady-state protocol to enhance long-term operation. Bioprocess Biosyst Eng 29:229–240PubMedCrossRefGoogle Scholar
  18. Parales RE, Haddock JD (2004) Biocatalytic degradation of pollutants. Curr Opin Biotechnol 15:374–379PubMedCrossRefGoogle Scholar
  19. Roch F, Alexander M (1997) Inability of bacteria to degrade low concentrations of toluene in water. Environ Toxicol Chem 16:1377–1383CrossRefGoogle Scholar
  20. Roy S, Gendron J, Delhomenie MC, Bibeau L, Heitz M, Brzezinski R (2003) Pseudomonas putida as the dominant toluene-degrading bacterial species during air decontamination by Biofiltration. Appl Microbiol Biotechnol 61:366–373PubMedGoogle Scholar
  21. Van Groenestijn JW, Hesselink PGM (1993) Biotechniques for air pollution control. Biodegradation 4:283–301CrossRefGoogle Scholar
  22. Williams TO, Miller FC (1992) Biofilters and facility operations. Biocycle 31:75–79Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Raúl Muñoz
    • 1
  • Luís Felipe Díaz
    • 1
  • Sergio Bordel
    • 1
  • Santiago Villaverde
    • 1
  1. 1.Department of Chemical Engineering and Environmental TechnologyUniversity of ValladolidValladolidSpain

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