Abstract
In this study the chlorophenol-degrading actinobacterium, Arthrobacter chlorophenolicus A6, was tested for its ability to grow on mixtures of phenolic compounds. During the experiments depletion of the compounds was monitored, as were cell growth and activity. Activity assays were based on bioluminescence output from a luciferase-tagged strain. When the cells were grown on a mixture of 4-chlorophenol, 4-nitrophenol and phenol, 4-chlorophenol degradation apparently was delayed until 4-nitrophenol was almost completely depleted. Phenol was degraded more slowly than the other compounds and not until 4-nitrophenol and 4-chlorophenol were depleted, despite this being the least toxic compound of the three. A similar order of degradation was observed in non-sterile soil slurries inoculated with A. chlorophenolicus. The kinetics of degradation of the substituted phenols suggest that the preferential order of their depletion could be due to their respective pKa values and that the dissociated phenolate ions are the substrates. A mutant strain (T99), with a disrupted hydroxyquinol dioxygenase gene in the previously described 4-chlorophenol degradation gene cluster, was also studied for its ability to grow on the different phenols. The mutant strain was able to grow on phenol, but not on either of the substituted phenols, suggesting a different catabolic pathway for the degradation of phenol by this microorganism.
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Abbreviations
- 4-BP:
-
4-bromophenol
- 4-CP:
-
4-chlorophenol
- 4-NP:
-
4-nitrophenol
References
Abuhamed T, Bayraktar E, Mehmetoglu T, Mehmetoglu U (2004) Kinetics model for growth of Pseudomonas putida F1 during benzene, toluene and phenol biodegradation. Process Biochem 39:983–988
Alexander M (1999) Biodegradation and bioremediation. Academic press, San Diego, CA
Alexander M, Lustigman BK (1966) Effect of chemical structure on microbial degradation of substituted benzenes. J Agric Food Chem 14:410–413
Backman A, Jansson JK (2004) Degradation of 4-chlorophenol at low temperature and during extreme temperature fluctuations by Arthrobacter chlorophenolicus A6. Microb Ecol 48:246–253
Bae HS, Lee JM, Kim YB, Lee ST (1996) Biodegradation of the mixtures of 4-chlorophenol and phenol by Comamonas testosteroni CPW301. Biodegradation 7:463–469
Baggi G, Cavalca L, Francia P, Zangrossi M (2004) Chlorophenol removal from soil suspensions: effects of a specialised microbial inoculum and a degradable analogue. Biodegradation 15:153–160
Cejkova A, Masak J, Jirku V, Vesely M, Patek M, Nesvera J (2005) Potential of Rhodococcus erythropolis as a bioremediation organism. World J Microbiol Biotechnol 21:317–321
Elväng AM, Westerberg K, Jernberg C, Jansson JK (2001) Use of green fluorescent protein and luciferase biomarkers to monitor survival and activity of Arthrobacter chlorophenolicus A6 cells during degradation of 4-chlorophenol in soil. Environ Microbiol 3:32–42
Escher BI, Snozzi M, Schwarzenbach RP (1996) Uptake, speciation, and uncoupling activity of substituted phenols in energy transducing membranes. Environ Sci Technol 30:3071–3079
Escher BI, Hunziker RW, Schwarzenbach RP (2001) Interaction of phenolic uncouplers in binary mixtures: concentration-additive and synergistic effects. Environ Sci Technol 35:3905–3914
Hao OJ, Kim MH, Seagren EA, Kim H (2002) Kinetics of phenol and chlorophenol utilization by Acinetobacter species. Chemosphere 46:797–807
Heinaru E, Viggor S, Vedler E, Truu J, Merimaa M, Heinaru A (2001) Reversible accumulation of p-hydroxybenzoate and catechol determines the sequential decomposition of phenolic compounds in mixed substrate cultivations in pseudomonads. FEMS Microbiol Ecol 37:79–89
Hollender J, Dott W, Hopp J (1994) Regulation of chloro- and methyl-phenol degradation in Comamonas testosteroni JH5. Appl Environ Microbiol 60:2330–2338
Jansson JK (2003) Marker and reporter genes: illuminating tools for environmental microbiologists. Curr Opin Biotech 6:310–316
Karigar C, Mahesh A, Nagenahalli M, Yun DJ (2006) Phenol degradation by immobilized cells of Arthrobacter citreus. Biodegradation 17:47–55
Kim JH, Oh KK, Lee ST, Kim SW, Hong SI (2002) Biodegradation of phenol and chlorophenols with defined mixed culture in shake-flasks and a packed bed reactor. Process Biochem 37:1367–1373
Kovárová-Kovar K, Egli T (1998) Growth kinetics of suspended microbial cells: from single-substrate-controlled growth to mixed-substrate kinetics. Microbiol Mol Biol R 62:646–666
Leung KT, Campbell S, Gan YD, White DC, Lee H, Trevors JT (1999) The role of the Sphingomonas species UG30 pentachlorophenol-4-monooxygenase in p-nitrophenol degradation. Fems Microbiol Lett 173:247–253
Lovanh N, Alvarez PJJ (2004) Effect of ethanol, acetate, and phenol on toluene degradation activity and tod-lux expression in Pseudomonas putida TOD102: Evaluation of the metabolic flux dilution model. Biotechnol Bioeng 86:801–808
McLaughlin SG, Dilger JP (1980) Transport of protons across membranes by weak acids. Physiol Rev 60:825–863
Nordin K, Unell M, Jansson JK (2005) Novel 4-chlorophenol degradation gene cluster and degradation route via hydroxyquinol in Arthrobacter chlorophenolicus A6. Appl Environ Microbiol 71:6538–6544
Reardon KF, Mosteller DC, Rogers JDB (2000) Biodegradation kinetics of benzene, toluene, and phenol as single and mixed substrates for Pseudomonas putida F1. Biotechnol Bioeng 69:385–400
Saez PB, Rittmann BE (1993) Biodegradation kinetics of a mixture containing a primary substrate (phenol) and an inhibitory co-metabolite (4-chlorophenol). Biodegradation 4:3–21
Sarand I, Skarfstad E, Forsman M, Romantschuk M, Shingler V (2001) Role of the DmpR-mediated regulatory circuit in bacterial biodegradation properties in methylphenol-amended soils. Appl Environ Microbiol 67:162–171
Stenberg B, Johansson M, Pell M, Sjodahl-Svensson K, Stenstrom J, Torstensson L (1998) Microbial biomass and activities in soil as affected by frozen and cold storage. Soil Biol Biochem 30:393–402
Stenström J (1989) Kinetics of decomposition of 2,4-dichlorophenoxyacetic acid by Alcaligenes eutrophus JMP134 and in soil. Toxicity Assessment 4:405–424
Terada H (1990) Uncouplers of oxidative phosphorylation. Environ Health Perspect 87:213–218
Travkin VM, Solyanikova IP, Golovleva LA (2006) Hydroxyquinol pathway for microbial degradation of halogenated aromatic compounds. J Environ Sci Health Part B-Pesticides Food Contaminants Agric Wastes 41:1361–1382
Unell M, Kabelitz N, Jansson JK, Heipieper HJ (2007) Adaptation of the psychrotroph Arthrobacter chlorophenolicus A6 to growth temperature and the presence of phenols by changes in the anteiso/iso ratio of branched fatty acids. Fems Microbiol Lett 266:138–143
Westerberg K, Elväng AM, Stackebrandt E, Jansson JK (2000) Arthrobacter chlorophenolicus sp nov, a new species capable of degrading high concentrations of 4-chlorophenol. Int J Syst Evol Microbiol 50:2083–2092
Xun LY, Topp E, Orser CS (1992) Diverse substrate range of a flavobacterium pentachlorophenol hydroxylase and reaction stoichiometries. J Bacteriol 174:2898–2902
Acknowledgements
The authors thank Therese Jernberg for excellent technical assistance. We also thank Karl-Heinz Gartemann and Rudolf Eichenlaub at Universität Bielefeld, Germany, for the kind gift of the pKGT452Cβ vector enabling us to create mutant T99. This work was supported by the Swedish Council for Engineering Science and the Swedish Foundation for Environmental Research (MISTRA).
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Unell, M., Nordin, K., Jernberg, C. et al. Degradation of mixtures of phenolic compounds by Arthrobacter chlorophenolicus A6. Biodegradation 19, 495–505 (2008). https://doi.org/10.1007/s10532-007-9154-2
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DOI: https://doi.org/10.1007/s10532-007-9154-2