Abstract
Phenol-degrading pseudomonads possessing different phenol hydroxylases (PH) were evaluated by the values of apparent half-saturation constant for phenol-oxygenating activity (K S ), maximum specific growth rate (μ max), lag-time length (λ), inhibition constant (K I ) and growth yield factor (Y X/S ). Strains of the same PH type showed similar kinetic parameters: single-component PH (sPH) harbouring strains had higher values of K S and lower values of μ max than the strains having multicomponent PH (mPH). However, the values of K I and the dependencies of the lag-time length on initial phenol concentration were strain-specific. The elevated ratio between specific activities of catechol 1,2-dioxygenase (C12O) and muconate cycloisomerase in sPH-strains caused irreversible accumulation of a high amount of exogenous cis,cis-muconate (CCM) which resulted in decreased Y X/S values. Co-presence of sPH and mPH genes did not give the strains PC16 and P69 any extra advantage and according to determined kinetic parameters only one PH was active during phenol degradation. At the same time simultaneous functioning of catechol ortho and meta cleavage pathways (strain PC20) resulted in higher μ max and Y X/S values. Evaluation of strains showed that the type of PH determined the efficiency of phenol degradation, whereas the tolerance to elevated phenol concentrations was strain-specific.
Similar content being viewed by others
References
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72(1–2):248–254
Cafaro V, Izzo V, Scognamiglio R, Notomista E, Capasso P, Casbarra A, Pucci P, Di Donato A (2004) Phenol hydroxylase and toluene/o-xylene monooxygenase from Pseudomonas stutzeri OX1: Interplay between two enzymes. Appl Environ Microbiol 70(4):2211–2219
Cafaro V, Notomista E, Capasso P, Di Donato A (2005) Regiospecificity of two multicomponent monooxygenases from Pseudomonas stutzeri OX1: Molecular basis for catabolic adaptation of this microorganism to methylated aromatic compounds. Appl Environ Microbiol 71(8):4736–4743
Chen W-M, Chang J-S, Wu C-H, Chang S-C (2004) Characterization of phenol and trichloroethene degradation by the rhizobium Ralstonia taiwanensis. Res Microbiol 155(8):672–680
Dagley S, Evans WC, Ribbons DW (1960) New pathways in the oxidative metabolism of aromatic compounds by micro-organsms. Nature 188:560–566
Ehrt S, Schirmer F, Hillen W (1995) Genetic organization, nucleotide sequence and regulation of expression of genes encoding phenol hydroxylase and catechol 1,2-dioxygenase in Acinetobacter calcoaceticus NCIB8250. Mol Microbiol 18(1):13–20
El Fantroussi S, Agathos SN (2005) Is bioaugmentation a feasible strategy for pollutant removal and site remediation? Curr Opin Microbiol 8:1–8
Ellison SLR, Rösslein M, Williams A (2000) Eurachem/CITAC guide, quantifying uncertainty in analytical measurement, 2nd edn. Eurachem, Berlin
Feist CF, Hegeman GD (1969) Phenol and benzoate metabolism by Pseudomonas putida: regulation of tangential pathways. J Bacteriol 100(2):869–877
Franck-Mokroβ AC, Schmidt E (1998) Simultaneous degradation of chloro- and methylsubstituted aromatic compounds: competition between Pseudomonas strains using the ortho and meta pathway or the ortho pathway exclusively. Appl Microbiol Biotechnol 50:233–240
Futamata H, Nagano Y, Watanabe K, Hiraishi A (2005) Unique kinetic properties of phenol-degrading Variovorax strains resposible for efficient trichloroethylene degradation in a chemostat enrichment culture. Appl Environ Microbiol 71(2):904–911
Goudar CT, Strevett KA (1998) Comparison of relative rates of BTEX biodegradation using respirometry. J Ind Microbiol Biotechnol 21:11–18
Healey FP (1980) Slope of the Monod equation as an indicator of advantage in nutrient competition. Microb Ecol 5(4):281–286
Hegeman GD (1966) Synthesis of the enzymes of the mandelate pathway by Pseudomonas putida. I. Synthesis of enzymes by the wild type. J Bacteriol 91(3):1140–1154
Heinaru E, Truu J, Stottmeister U, Heinaru A (2000) Three types of phenol and p-cresol-degrading bacteria isolated from river water continuously polluted with phenolic compounds. FEMS Microbiol Ecol 31:195–205
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
Heinaru E, Merimaa M, Viggor S, Lehiste M, Leito I, Truu J, Heinaru A (2005) Biodegradation efficiency of functionally important population selected for bioaugmentation in phenol- and oil-polluted area. FEMS Microbiol Ecol 51:363–373
Hino S, Watanabe K, Takahashi N (1998) Phenol hydroxylase cloned from Ralstonia eutropha strain E2 exhibits novel kinetic properties. Microbiology 144:1765–1772
Jiang H-L, Tay ST-L, Maszenan AM, Tay J-H (2006) Physiological traits of bacterial strains isolated from phenol-degrading aerobic granules. FEMS Microbiol Ecol 57:182–191
Kivisaar M, Horak R, Kasak L, Heinaru A, Habicht J (1990) Selection of independent plasmids determining phenol degradation in Pseudomonas putida and the cloning and expression of genes encoding phenol monooxygenase and catechol 1,2-dioxygenase. Plasmid 24(1):25–36
Kivisaar M, Kasak L, Nurk A (1991) Sequence of the plasmid-encoded catechol 1,2-dioxygenase-expressing gene, pheB, of phenol-degrading Pseudomonas sp. strain EST1001. Gene 98(1):15–20
Kiesel B, Müller RH (2002) The meta pathway as a potential energy-generating secuence and its effects on the growth rate during the utilisation of aromatics. Acta Biotechol 22(3–4):221–134
Kovárová- Kovar K, Egli T (1998) Growth kinetics of suspended microbial cells: from single-substrate-controlled growth to mixed-substrate kinetics. Microbiol Mol Biol Rev 62(3):646–666
Kukor JJ, Olsen RH (1990) Molecular cloning, characterization, and regulation of a Pseudomonas pickettii PKO1 gene encoding phenol hydroxylase and expression of the gene in Pseudomonas aeruginosa PAO1c. J Bacteriol 172:4624–4630
Kukor JJ, Olsen RH (1991) Genetic organization and regulation of a meta cleavage pathway for catechols produced from catabolism of toluene, benzene, phenol and cresols by Pseudomonas pickettii PKO1. J Bacteriol 173(15):4587–4594
Merimaa M, Heinaru E, Liivak M, Vedler E, Heinaru A (2006) Grouping of phenol hydroxylase and catechol 2,3-dioxygenase genes among phenol- and p-cresol-degrading Pseudomonas species and biotypes. Arch Microbiol 186:287–296
Müller RH, Babel W (1996) Growth rate-dependent expression of phenol-assimilation pathways in Alcaligenes eutrophus JMP 134—the influence of formate as an auxiliary energy source on phenol conversion characteristics. Appl Microbiol Biotechnol 46:156–162
Ng LC, Shingler V, Sze CC, Poh CL (1994) Cloning and sequences of the first eight genes of the chromosomally encoded (methyl) phenol degradation pathway from Pseudomonas putida P35X. Gene 151(1–2):29–36
Nurk A, Kasak L, Kivisaar M (1991) Sequence of the gene (pheA) encoding phenol monooxygenase from Pseudomonas sp. EST1001: expression in Escherichia coli and Pseudomonas putida. Gene 102(1):13–18
Ornston LN (1971) Regulation of catabolic pathways in Pseudomonas. Bacteriol Rev 35(2):87–116
Ornston LN, Stanier RY (1966) The conversion of catechol and protocatechuate to beta-ketoadipate by Pseudomonas putida. I. Biochemistry. J Biol Chem 241(16):3776–3786
Peters M, Heinaru E, Talpsep E, Wand H, Stottmeister U, Heinaru A, Nurk A (1997) Acquisition of a deliberately introduced phenol degradation operon, pheBA, by different indigenous Pseudomonas species. Appl Environ Microbiol 63(12):4899–4906
Pessione E, Divari S, Griva E, Cavaletto M, Rossi GL, Gilardi G, Gilunta C (1999) Phenol hydroxylase from Acinetobacter radioresistens is a multicomponent enzyme. Purification and characterization of the reductase moiety. Eur J Biochem 265(2):549–555
Pieper DH, Stadler-Fritzsche K, Knackmuss H-J, Timmis KN (1995) Formation of dimethylmuconolactones from dimethylphenols by Alcaligenes eutrophus JMP 134. Appl Environ Microbiol 61(6):2159–2165
Polymenakou PN, Stephanou EG (2005) Effect of temperature and additional carbon sources on phenol degradation by an indigenous soil Pseudomonad. Biodegradation 16:403–413
Shingler V, Franklin CH, Tsuda M, Holroyd D, Bagdasarian M (1989) Molecular analysis of a plasmid-encoded phenol hydroxylase from Pseudomonas CF600. J Gen Microbiol 135:1083–1092
Takeo M, Maeda Y, Okada H, Miyama K, Mori K, Ike M, Fujita M (1995) Molecular cloning and sequencing of the phenol hydroxylase gene from Pseudomonas putida BH. J Ferment Bioeng 79(5):485–488
Vollmer MD, Hoier H, Hecht H-J, Schell U, Gröning J, Goldman A, Schlömann M (1998) Substrate specificity of and product formation by muconate cycloisomerases: an analysis of wild-type enzymes and engineered variants. Appl Environ Microbiol 64(9):3290–3299
Watanabe K, Teramoto M, Futamata H, Harayama S (1998) Molecular detection, isolation, and physiological characterization of functionally dominant phenol-degrading bacteria in activated sludge. Appl Environ Microbiol 64(11):4396–4402
Watanabe K, Futamata H, Harayama S (2002) Understanding the diversity in catabolic potential of microorganisms for the development of bioremediation strategies. Antonie Van Leeuwenhoek 81:655–663
Whiteley AS, Wiles S, Lilley AK, Philp J, Bailey MJ (2001) Ecological and physiological analyses of Pseudomonad species within a phenol remediation system. J Microbiol Methods 44:79–88
Williams PA, Shaw LE (1997) mucK, a gene in Acinetobacter calcoaceticus ADP1 (BD413), encodes the ability to grow on exogenous cis,cis-muconate as the sole carbon source. J Bacteriol 179(18):5935–5942
Zwietering MH, Jongenburger I, Rombouts FM, van´t Riet K (1990) Modeling of the bacterial growth curve. Appl Environ Microbiol 56(6):1875–1881
Acknowledgments
The Estonian Science Foundation, Grant 5682, and the Institute of Molecular and Cell Biology, University of Tartu, supported this research. We gratefully acknowledge Prof. I. Leito, Dr. T. Alamäe and Dr. K. Orupõld for helpful discussions.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Viggor, S., Heinaru, E., Künnapas, A. et al. Evaluation of different phenol hydroxylase-possessing phenol-degrading pseudomonads by kinetic parameters. Biodegradation 19, 759–769 (2008). https://doi.org/10.1007/s10532-008-9180-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10532-008-9180-8